Histone deacetylase inhibitors and methods of use thereof

ABSTRACT

The invention provides novel classes of HDAC inhibitors. Methods of sensitizing a cancer cell to the cytotoxic effects of radiotherapy are also provided as well as methods for treating cancer and methods for treating neurological diseases. Additionally, the invention further provides pharmaceutical compositions comprising an HDAC inhibitor of the invention, and kits comprising a container containing an HDAC inhibitor of the invention.

This application is a continuation of U.S. patent application Ser. No.10/614,498, filed Jul. 7, 2003, now U.S. Pat. No. 7,842,835.

1. FIELD OF THE INVENTION

The present invention relates to histone deacetylase (“HDAC”)inhibitors, pharmaceutical compositions comprising an HDAC inhibitor,methods of increasing the sensitivity of a cancer cell to the cytotoxiceffects of radiotherapy comprising contacting said cell with an HDACinhibitor, and methods of treating cancer or a neurological diseasecomprising administering to a subject in need thereof, an HDACinhibitor.

2. BACKGROUND OF THE INVENTION 2.1 Cancer

Cancer is the second leading cause of death in the United States afterheart disease. The American Cancer Society estimated that in 2002, therewere 1.3 million new cases of cancer and 555,000 cancer-related deaths.Overall mortality rates have declined by 1% per year during the 1990s.There are currently over 9 million living Americans who have beendiagnosed with cancer and the NIH estimates the direct medical costs ofcancer as $60 billion per year.

Typical treatment modalities useful in the treatment of cancer includechemotherapy, radiotherapy and surgery (see, for example, Stockdale,1998, “Principles of Cancer Subject Management”, in Scientific American:Medicine, vol. 3, Rubenstein and Federman, eds., Chapter 12, SectionIV). All of these approaches pose significant drawbacks for the subject.Surgery, for example, can be contraindicated due to the health of thesubject or can be unacceptable to the subject. Additionally, surgery maynot successfully remove all neoplastic tissue. Chemotherapy involves theadministration of cytotoxic chemical agents which are associated with abroad spectrum of undesirable side effects, including alopecia, nauseaand vomiting, hematoxicity, neurotoxicity, nephrotoxicity,cardiotoxicity and hepatotoxicity. In addition, cancer cells commonlydevelop resistance to most anticancer agents, thus renderingchemotherapy ineffective over time.

Radiation therapy, or radiotherapy as it is sometimes referred to,involves the treatment of cancer and other diseases using ionizingradiation. Ionizing radiation deposits energy that injures or destroyscells in targeted tissues by damaging their genetic material andsubsequently interfering with a cell's ability to grow and/or replicate.Although radiation causes damage to both cancer cells and normal cells,the latter are better able to repair themselves and continue to functionproperly. Radiotherapy can be used to treat localized solid tumors, suchas cancers of the skin, tongue, larynx, brain, breast, prostate, colon,uterus, lung, kidney, head and neck, and/or cervix. It can also be usedto treat systemic forms of cancer such as the leukemias and lymphomas.

Radiotherapy is optimally effective when the targeted neoplastic tissueexhibits a higher sensitivity to the effects of radiation thanneighboring normal tissue. In the absence of such differences insensitivity, radiotherapy often elicits serious side effects.

Radiation responses of tumors vary as a function of histology, doublingtime, oxygenation, availability of nutrients, repair capacity and otherfactors. Peters et al., Int J. Radiat. Biol., 1994, 66:523-529. Certaintypes of cancer are readily cured using ionizing radiation doses withinnormal tissue tolerances, while other types of cancer are not veryresponsive to radiation. Furthermore, radiation responses of tumors withthe same histology may show considerable heterogeneity and reduce thetherapeutic effects of the therapy. Weichselbaum et al, Int. J. Radiat.Oncol. Biol. Phys., 1988, 14:907-912. Thus, a primary challenge facingradiotherapy is the differentiation between the more radiosensitivetumors vs. less radiosensitive tumors.

Investigations into the molecular bases underlying cellular radiationresponses have provided dramatic mechanistic insight. Signaltransduction pathways have been implicated to play important roles incellular responses to ionizing radiation. Kornberg et al., Twenty-fiveyears of the Nucleosome, Fundamental Particle of the EukaryoteChromosome, Cell Press 1999, 98:285-294. Induction of gene expression bythese cascades under various conditions has been shown to result in cellcycle arrest, activation of DNA repair processes, and activation ofprogrammed cell death (apoptosis). Meyn, Cancer Res., 1995,55:5991-6001, and Jackson et al., Trends Biochem. Sci., 1995,20:412-415. Disruption of critical signaling pathways in cancer cellsresults in enhanced cytotoxic effects following radiation exposure.

Histone acetylation and deacetylation play important roles in chromatinfolding and maintenance. Kornberg et al., Bjorklund et al., Cell, 1999,96:759-767, and Struhl et al., Cell, 1998, 94:1-4. Acetylated chromatinis more open and has been implicated in the increased radiationsensitivities observed in some cell types. Oleinick et al., Int. J.Radiat. Biol., 1994, 66:523-529. Furthermore, certainradiation-resistant human cancer cells treated with the histonedeacetylase (HDAC) inhibitor, trichostatin A (TSA), were sensitized tothe damaging effects of ionizing radiation. Thus, HDAC inhibitors may beuseful as radiation sensitizing agents.

There is a significant need in the art for novel compounds,compositions, and methods that are useful for treating cancer orneoplastic disease with increased selectivity and decreased toxicity.

2.2 Neurological Diseases

Millions of people worldwide suffer from debilitating neurologicaldiseases. Neurological diseases affect a vast number of humans of allages (see Table 328-2 In: Wyngaarden and Smith, 1988, Cecil Textbook ofMedicine, 18^(th) Ed., W.B. Saunders Co., Philadelphia, pp. 1750-1753).Each year in the United States alone, over 500,000 people experience astroke, making it the third leading cause of death and the primary causeof disabililty. One in twenty people is afflicted with Alzheimer'sdisease by the age of 65, and almost 40 percent of the population havethe disease by age 80. More than 600,000 people suffer from Parkinson'sdisease and over 200,000 from multiple sclerosis. Every year, greaterthan 10,000 people die from amyotrophic lateral sclerosis (ALS). Theimpact of neurological disease is not only devastating not only forpatients, but also for their families

Although considerable effort has been invested in the design ofeffective therapies, neurological diseases continue to threaten andlessen the qualitity of the lives of millions of people worldwide.

Accordingly, there is a need in the art for improved compounds,compositions, and methods useful for the treatment of neurologicaldiseases.

The recitation of any reference in Section 2 of this application is notan admission that the reference is prior art to this application.

3. SUMMARY OF THE INVENTION

The present invention encompasses HDAC inhibitors, pharmaceuticalcompositions compositions comprising an HDAC inhibitor, and methods fortreating cancer or a neurological disease comprising administering anHDAC inhibitor to a subject in need thereof.

Accordingly, in one embodiment, the invention provides compounds havingthe Formula (I):

-   -   and pharmaceutically acceptable salts thereof,    -   wherein    -   R¹ is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more -halo, —C₁-C₆ alkyl, —O—(C₁-C₆        alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or        —C(O)NHR′ groups wherein R′ is —H or unsubstituted —C₁-C₆ alkyl;        with the proviso that when n is 2, R¹ cannot be —C₃-C₇        cycloalkyl or -3- to 10-membered heterocycle;    -   m is an integer ranging from 1-10; and    -   n is an integer ranging from 1-10.

In another embodiment, the invention further provides compounds havingthe Formula (Ia):

and pharmaceutically acceptable salts thereof,

wherein

-   -   R^(1a) is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more -halo, —C₁-C₆ alkyl, —O—(C₁-C₆        alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or        —C(O)NHR′ groups wherein R′ is —H or unsubstituted —C₁-C₆ alkyl;    -   m is an integer ranging from 0-10; and    -   n is an integer ranging from 1-10.

In a further embodiment, the invention further provides compounds havingthe Formula (II):

and pharmaceutically acceptable salts thereof,

wherein

-   -   Y is —C(O)CH₂SH or —NHC(O)CH₂SH;    -   R² is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more -halo, —C₁-C₆ alkyl, —O—(C₁-C₆        alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or        —C(O)NHR′ groups wherein R′ is —H or unsubstituted —C₁-C₆ alkyl;    -   m is an integer ranging from 0-10; and    -   n is an integer ranging from 1-10.

In still another embodiment, the invention further provides compoundshaving the Formula (III):

and pharmaceutically acceptable salts thereof,

wherein

-   -   Z is —C(O)NHOH, —C(O)CH₂SH or —NHC(O)CH₂SH;    -   R³ is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl, -3- to 10-membered        heterocycle, any of which may be unsubstituted or substituted        with one or more -halo, —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH,        —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′        groups wherein R′ is —H or unsubstituted —C₁-C₆ alkyl;    -   R⁴ is —H or —Si(R⁵)₃;    -   each occurrence of R⁵ is independently unsubstituted —C₁-C₆        alkyl;    -   m is an integer ranging from 0-10; and    -   n is an integer ranging from 1-10.

In yet another embodiment, the invention further provides compoundshaving the Formula (IV):

and pharmaceutically acceptable salts thereof,

wherein

-   -   R⁶ is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more -halo, —C₁-C₆ alkyl, —O—(C₁-C₆        alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or        —C(O)NHR′ groups wherein R′ is —H or unsubstituted —C₁-C₆ alkyl;    -   m is 1 or an integer ranging from 8-10; and    -   n is an integer ranging from 1-10.

In another embodiment, the invention further provides compounds havingthe Formula (IVa):

and pharmaceutically acceptable salts thereof,

wherein

-   -   R^(6a) is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more -halo, —C₁-C₆ alkyl, —O—(C₁-C₆        alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or        —C(O)NHR′ groups wherein R′ is —H or unsubstituted —C₁-C₆ alkyl;    -   m is an integer ranging from 0-10; and    -   n is an integer ranging from 2-10.

In a further embodiment, the invention further provides compounds havingthe Formula (V):

and pharmaceutically acceptable salts thereof,

wherein

-   -   Y is —C(O)CH₂SH or —NHC(O)CH₂SH;    -   R⁷ is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more -halo, —C₁-C₆ alkyl, —O—(C₁-C₆        alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or        —C(O)NHR′ groups wherein R′ is —H or unsubstituted —C₁-C₆ alkyl;        with the proviso that when n is 2, R⁷ cannot be —C₃-C₇        cycloalkyl or -3- to 10-membered heterocycle;    -   m is an integer ranging from 0-10; and    -   n is an integer ranging from 1-10.

In another embodiment, the invention further provides compounds havingthe Formula (VI):

and pharmaceutically acceptable salts thereof,

wherein

-   -   each Z is independently —C(O)NHOH, —C(O)CH₂SH or —NHC(O)CH₂SH,        with the proviso that when both Z groups are —C(O)NHOH, the        phenyl group of said compound of formula (VI) is either ortho or        meta substituted;    -   m is an integer ranging from 1-10; and    -   n is an integer ranging from 1-10.

In yet another embodiment, the invention further provides compoundshaving the Formula (VII):

and pharmaceutically acceptable salts thereof,

wherein

-   -   each Y is independently —C(O)CH₂SH or —NHC(O)CH₂SH;    -   m is an integer ranging from 1-10; and    -   n is an integer ranging from 1-10.

In a further embodiment, the invention further provides compounds havingthe Formula (VIII):

and pharmaceutically acceptable salts thereof,

wherein:

-   -   each R⁸ is independently —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl        or -3- to 10-membered heterocycle, any of which may be        unsubstituted or substituted with one or more -halo,        —C₁-C₆alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′,        N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or        unsubstituted —C₁-C₆alkyl;    -   each G is independently —NH— or —CH₂—;    -   each J is independently —NH— or —CH₂—;    -   each m is independently an integer ranging from 1-10; and    -   each n is independently an integer ranging from 1-10.

In a further embodiment, the invention further provides compounds havingthe Formula (IX):

and pharmaceutically acceptable salts thereof,

wherein

-   -   R⁹ is phenyl, which can be unsubstituted or substituted with one        or more -halo, —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′,        —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′        is —H or unsubstituted —C₁-C₆ alkyl; and    -   m is an integer ranging from 2-10.

The invention also provides pharmaceutical compositions comprising thecompound of Formula (I), (II), (III), (IV), (V), (VI), (VII), (VIII) or(IX), or a pharmaceutically acceptable salt thereof, in an amountsufficient to treat said cancer, and a pharmaceutically acceptablecarrier or vehicle. The compositions are useful for treating cancer.

The present invention further provides methods for increasing thesensitivity of a cancer cell to the cytotoxic effects of radiotherapy,said method comprising contacting said cell with the compound orpharmaceutically acceptable salt of the compound of Formula (I), (Ia),(II), (III), (IV), (IVa), (V), (VI), (VII), (VIII) or (IX), in an amountsufficient to increase the sensitivity of said cell to the cytotoxiceffects of radiotherapy.

The invention also provides methods for treating cancer, said methodscomprising administering to a subject in need thereof the compound orpharmaceutically acceptable salt of the compound of Formula (I), (II),(III), (IV), (V), (VI), (VII), (VIII) or (IX), in an amount sufficientto treat said cancer.

The invention further provides methods for treating cancer, said methodscomprising:

(a) administering to a subject in need thereof, the compound orpharmaceutically acceptable salt of the compound of Formula (I), (Ia),(II), (III), (IV), (IVa), (V), (VI), (VII), (VIII) or (IX), in an amountsufficient to sensitize a cancer cell to the cytotoxic effects ofradiotherapy; and

(b) administering to said subject an amount of radiotherapy sufficientto treat said cancer.

The present invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound or pharmaceutically acceptable salt of the compoundof Formula (I), (Ia), (II), (III), (IV), (IVa), (V), (VI), (VII), (VIII)or (IX), in an amount sufficient to treat said neurological disease.

The present invention also provides kits comprising a container whichcontains the compound or pharmaceutically acceptable salt of thecompound of Formula (I), (Ia), (II), (III), (IV), (IVa), (V), (VI),(VII), (VIII) or (IX).

The details of the invention are set forth in the accompanyingdescription below. Although any methods and materials similar orequivalent to those described herein can be used in the practice ortesting of the present invention, illustrative methods and materials arenow described. Other features, objects, and advantages of the inventionwill be apparent from the description and from the claims. In thespecification and the appended claims, the singular forms also includethe plural unless the context clearly dictates otherwise. Unless definedotherwise, all technical and scientific terms used herein have the samemeaning as commonly understood by one of ordinary skill in the art towhich this invention belongs. All patents, patent applications andpublications cited in this specification are incorporated herein byreference for all purposes.

4. BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates the inhibitory effect of selected compounds of theinvention on HDAC activity in HeLa nuclear cell extracts. Data isexpressed as arbitrary fluorescence units (AFU)/μM obtained with theobserved range of values obtained in the enzyme assays used in a seriesof dilutions for a standard curve. Data is shown for a blank sample (noenzyme), a control sample (no inhibitor), the known compound MD83A (as anegative control) at 3 μM, the known HDAC inhibitor TSA at 0.5 μM and 5μM, the known HDAC inhibitor SAHA at 1 μM and 5 μM, and Compounds of theInvention 2, 3, 5, 6, 7, 8, 9, 10, 11, 21, 22, 23, 24 and 25, each at 5μM.

5. DETAILED DESCRIPTION OF THE INVENTION 5.1 Definitions

The term “C₁-C₆ alkyl” as used herein refers to a straight or branchedchain, saturated or unsaturated hydrocarbon having from 1 to 6 carbonatoms. Representative C₁-C₆ alkyl groups include, but are not limited tomethyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-buty, pentyl,isopentyl, neopentyl, hexyl, isohexyl, neohexyl, ethylenyl, propylenyl,1-butenyl, 2-butenyl, 1-pentenyl, 2-pentenyl, 1-hexenyl, 2-hexenyl,3-hexenyl, acetylenyl, pentynyl, 1-butynyl, 2-butynyl, 1-pentynyl,2-pentynyl, 1-hexynyl, 2-hexynyl and 3-hexynyl. A C₁-C₆ alkyl group maybe unsubstituted or substituted with one or more of the followinggroups: -halo, —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′,—OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —Hor unsubstituted —C₁-C₆ alkyl.

The term “aryl” as used herein refers to a phenyl group or a naphthylgroup. An aryl group may be unsubstituted or substituted with one ormore of the following groups: -halo, —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl),—OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groupswherein R′ is —H or unsubstituted —C₁-C₆ alkyl.

The phrase “Compounds of the Invention” as used herein refers to acompound of Formula (I), (Ia), (II), (III), (IV), (IVa), (V), (VI),(VII), (VIII) or (IX) or a pharmaceutically acceptable salt thereof. Insome instances, it is possible for a Compound of the Invention to haveone or more chiral centers. In these instances, it is to be understoodthat the invention encompasses all possible stereoisomers of thesecompounds.

The term “C₃-C₇ cycloalkyl” as used herein is a 3-, 4-5-, 6- or7-membered saturated or unsaturated non-aromatic carbocyclic ring.Representative C₃-C₇ cycloalkyls include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclopentadienyl, cyclohexyl,cyclohexenyl, cycloheptyl, cycloheptanyl, 1,3-cyclohexadienyl,-1,4-cyclohexadienyl, -1,3-cycloheptadienyl, and-1,3,5-cycloheptatrienyl. A C₃-C₇ cycloalkyl group may be unsubstitutedor substituted with one or more of the following groups: -halo, —C₁-C₆alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂,—NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or unsubstituted —C₁-C₆alkyl.

The term “halo” as used herein refers to —F, —Cl, —Br or —I.

As used herein, a “-3- to 10-membered heterocycle” is a 3- to10-membered aromatic or nonaromatic monocyclic or bicyclic ring ofcarbon atoms and from 1 to 4 heteroatoms selected from oxygen, nitrogenand sulfur. Examples of 3- to 10-membered heterocycles include, but arenot limited to, aziridinyl, oxiranyl, thiiranyl, azirinyl, diaziridinyl,diazirinyl, oxaziridinyl, azetidinyl, azetidinonyl, oxetanyl, thietanyl,piperidinyl, piperazinyl, morpholinyl, pyrrolyl, oxazinyl, thiazinyl,diazinyl, triazinyl, tetrazinyl, imidazolyl, benzimidazolyl, tetrazolyl,indolyl, isoquinolinyl, quinolinyl, quinazolinyl, pyrrolidinyl, purinyl,isoxazolyl, benzisoxazolyl, furanyl, furazanyl, pyridinyl, oxazolyl,benzoxazolyl, thiazolyl, benzthiazolyl, thiophenyl, pyrazolyl,triazolyl, benzodiazolyl, benzotriazolyl, pyrimidinyl, isoindolyl andindazolyl. A -3- to 10-membered heterocycle group may be unsubstitutedor substituted with one or more of the following groups: -halo, —C₁-C₆alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂,—NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or unsubstituted —C₁-C₆alkyl.

The Compounds of the Invention can be formulated as pharmaceuticallyacceptable salts. The phrase “pharmaceutically acceptable salt,” as usedherein, refers to a pharmaceutically acceptable organic or inorganicacid or base salt of an organic chemical compound. Representative“pharmaceutically acceptable salts” include, e.g., water-soluble andwater-insoluble salts, such as the acetate, amsonate(4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzonate,bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium,calcium edetate, camsylate, carbonate, chloride, citrate, clavulariate,dihydrochloride, edetate, edisylate, estolate, esylate, fiunarate,gluceptate, gluconate, glutamate, glycollylarsanilate,hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide,hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate,lactobionate, laurate, malate, maleate, mandelate, mesylate,methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate,N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate,oxalate, palmitate, pamoate (1,1-methene-bis-2-hydroxy-3-naphthoate,einbonate), pantothenate, phosphate/diphosphate, picrate,polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate,subacetate, succinate, sulfate, sulfosaliculate, suramate, tannate,tartrate, teoclate, tosylate, triethiodide, and valerate salts. Thecounterion may be any organic or inorganic moiety that stabilizes thecharge on the parent compound. Furthermore, a pharmaceuticallyacceptable salt may have more than one charged atom in its structure. Inthis instance the pharmaceutically acceptable salt can have multiplecounterions. Hence, a pharmaceutically acceptable salt can have one ormore charged atoms and/or one or more counterions.

As used herein, the term “purified” means that when isolated (e.g., fromother components of a synthetic organic chemical reaction mixture), theisolate contains at least 30%, at least 35%, at least 40%, at least 45%,at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95% or atleast 98% of a Compound of the Invention by weight of the isolate. In apreferred embodiment, the isolate contains at least 95% of a Compound ofthe Invention by weight of the isolate.

The following abbreviations are used herein and have the indicateddefinitions:

DMSO is dimethylsulfoxide, DTT is dithiothreitol, EDCI is1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, EDTA isethylenediaminetetraacetic acid, Et₃N is triethylamine, EtOAc is ethylacetate, HDAC is histone deacetylase, HEPES isN-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid, MeOH is methanol,MS is mass spectrometry, NMR is nuclear magnetic resonance, PBS isphosphate buffered saline, PDV is, SAHA is suberoylanilide hydroxamicacid, TBS is tert-butyldimethylsilyl, THF is tetrahydrofuran, Tr istrityl(triphenylmethyl), and TSA is trichostatin A(7-(4-(dimethylamino)phenyl)-n-hydroxy-4,6-dimethyl-7-oxo-2,4-heptadienamide).

5.2 The Compounds of the Invention 5.2.1 The Compounds of Formula (I)

As stated above, the present invention encompasses compounds having theFormula (I)

and pharmaceutically acceptable salts thereof,

wherein

-   -   R′ is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more of the following groups: -halo,        —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′,        NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or        unsubstituted —C₁-C₆ alkyl; with the proviso that when n is 2,        R¹ cannot be —C₃-C₇ cycloalkyl or -3- to 10-membered        heterocycle;    -   m is an integer ranging from 1-10; and    -   n is an integer ranging from 1-10.

A first subclass of the compounds of Formula (I) is that wherein R¹ isphenyl.

A second subclass of the compounds of Formula (I) is that wherein n isan integer ranging from 1 to 5.

A third subclass of the compounds of Formula (I) is that wherein m is 2.

A fourth subclass of the compounds of Formula (I) is that wherein m is 1and R¹ is 4-(N,N-dimethylamino)phenyl.

Illustrative Compounds of Formula (I) include the compounds listedbelow:

Compound No. R¹ m n 1 Phenyl 2 3 2 4-N(CH₃)₂-Phenyl 1 3 34-N(CH₃)₂-Phenyl 1 4 4 4-N(CH₃)₂-Phenyl 1 5 5 4-N(CH₃)₂-Phenyl 1 6 64-N(CH₃)₂-Phenyl 1 7and pharmaceutically acceptable salts thereof.

The present invention also provides pharmaceutical compositionscomprising the compound of Formula (I) or a pharmaceutically acceptablesalt thereof, and a pharmaceutically acceptable carrier or vehicle.

The invention also provides a method for treating cancer, said methodcomprising administering to a subject in need thereof the compound ofFormula (I) or a pharmaceutically acceptable salt thereof, in an amountsufficient to treat said cancer.

The invention further provides method for increasing the sensitivity ofa cancer cell to the cytotoxic effects of radiotherapy, said methodcomprising contacting said cell with the compound of Formula (I) in anamount sufficient to increase the sensitivity of said cell to thecytotoxic effects of radiotherapy.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (I) or a pharmaceutically acceptable salt thereof; in an        amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound of Formula (I), or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

5.2.2 The Compounds of Formula (Ia)

As stated above, the present invention encompasses compounds having theFormula (Ia):

and pharmaceutically acceptable salts thereof,

wherein

-   -   R′ is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more of the following groups: -halo,        —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′,        NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or        unsubstituted —C₁-C₆ alkyl;    -   m is an integer ranging from 0-10; and    -   n is an integer ranging from 1-10.

Illustrative examples of compounds of Formula (Ia) include the compoundslisted below:

Compound No. R¹ m n 7 4-N(CH₃)₂-Phenyl 0 6 8 Adamantyl 0 5and pharmaceutically acceptable salts thereof.

The invention further provides a method for increasing the sensitivityof a cancer cell to the cytotoxic effects of radiotherapy, said methodcomprising contacting said cell with the compound of Formula (Ia) or apharmaceutically acceptable salt thereof, in an amount sufficient toincrease the sensitivity of said cell to the cytotoxic effects ofradiotherapy.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (Ia) or a pharmaceutically acceptable salt thereof, in        an amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof, the compound of Formula (Ia) or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

5.2.3 The Compounds of Formula (II)

As stated above, the present invention encompasses compounds having theFormula (II)

and pharmaceutically acceptable salts thereof,

wherein

-   -   Y is —C(O)CH₂SH or —NHC(O)CH₂SH;    -   R² is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more of the following groups: -halo,        —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′,        NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or        unsubstituted —C₁-C₆ alkyl;    -   m is an integer ranging from 0-10; and    -   n is an integer ranging from 1-10.

A first subclass of the compounds of Formula (II) is that wherein m is1.

A second subclass of the compounds of Formula (II) is that wherein R² is4-(N,N-dimethylamino)phenyl.

A third subclass of the compounds of Formula (II) is that wherein m is 1and R² is 4-(N,N-dimethylamino)phenyl.

Illustrative Compounds of Formula (II) include the compounds listedbelow:

Compound No. R² Y m n 9 Phenyl —NHC(O)CH₂SH 0 5 10 Phenyl —NHC(O)CH₂SH 06 11 Phenyl —NHC(O)CH₂SH 1 5 12 4-N(CH₃)₂-Phenyl —NHC(O)CH₂SH 1 6 13Phenyl —NHC(O)CH₂SH 0 6and pharmaceutically acceptable salts thereof.

The present invention also provides pharmaceutical compositionscomprising the compound of Formula (II) or a pharmaceutically acceptablesalt thereof, and a pharmaceutically acceptable carrier or vehicle.

The invention also provides a method for increasing the sensitivity of acancer cell to the cytotoxic effects of radiotherapy, said methodcomprising contacting said cell with the compound or a pharmaceuticallyacceptable salt of the compound of Formula (II) effective to increasethe sensitivity of said cell to the cytotoxic effects of radiotherapy.

The invention also provides a method for treating cancer, said methodcomprising administering to a subject in need thereof the compound ofFormula (II) or a pharmaceutically acceptable salt thereof, in an amountsufficient to treat said cancer.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (II) or a pharmaceutically acceptable salt thereof, in        an amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound of Formula (II) or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

5.2.4 The Compounds of Formula (III)

As stated above, the present invention encompasses compounds having theFormula (III)

and pharmaceutically acceptable salts thereof,

wherein

-   -   Z is —C(O)NHOH, —C(O)CH₂SH or —NHC(O)CH₂SH;    -   R³ is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl, -3- to 10-membered        heterocycle, any of which may be unsubstituted or substituted        with one or more of the following groups: -halo, —C₁-C₆ alkyl,        —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂,        —NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or unsubstituted        —C₁-C₆ alkyl;    -   R⁴ is —H or —Si(R⁵)₃;    -   each occurrence of R⁵ is independently unsubstituted —C₁-C₆        alkyl;    -   m is an integer ranging from 0-10; and    -   n is an integer ranging from 1-10.

A first subclass of the compounds of Formula (III) is that wherein m is2.

A second subclass of the compounds of Formula (III) is that wherein n is2 or 3.

A third subclass of the compounds of Formula (III) is that wherein R⁴ is—H.

A fourth subclass of the compounds of Formula (III) is that wherein R³is phenyl.

Illustrative examples of Compounds of Formula (III) include thecompounds listed below:

Compound No. R³ R⁴ Z m n 14 Phenyl H —C(O)NHOH 1 2 15 Phenyl H —C(O)NHOH1 3 16 Phenyl —Si(CH₃)₂(t-butyl) —C(O)NHOH 1 3and pharmaceutically acceptable salts thereof.

The present invention also provides pharmaceutical compositionscomprising the compound of Formula (III) or a pharmaceuticallyacceptable salt thereof; and a pharmaceutically acceptable carrier orvehicle.

The invention also provides a method for increasing the sensitivity of acancer cell to the cytotoxic effects of radiotherapy, said methodcomprising contacting said cell with the compound of Formula (III) or apharmaceutically salt thereof, in an amount sufficient to increase thesensitivity of said cell to the cytotoxic effects of radiotherapy.

The invention also provides a method for treating cancer, said methodcomprising administering to a subject in need thereof the compound ofFormula (III) or a pharmaceutically acceptable salt thereof, in anamount sufficient to treat said cancer.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (III) or a pharmaceutically acceptable salt thereof, in        an amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound of Formula (III) or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

5.2.5 The Compounds of Formula (IV)

As stated above, the present invention encompasses compounds having theFormula (IV):

and pharmaceutically acceptable salts thereof,

wherein

-   -   R⁶ is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more of the following groups: -halo,        —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′,        NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or        unsubstituted —C₁-C₆ alkyl;    -   m is 1 or an integer ranging from 8-10; and    -   n is an integer ranging from 1-10.

The present invention also provides pharmaceutical compositionscomprising a compound of Formula (IV) or a pharmaceutically acceptablesalt thereof, and a pharmaceutically acceptable carrier or vehicle.

A method for increasing the sensitivity of a cancer cell to thecytotoxic effects of radiotherapy, said method comprising contactingsaid cell with the compound of Formula (IV) or pharmaceutically saltthereof, effective to increase the sensitivity of said cell to thecytotoxic effects of radiotherapy.

The invention also provides a method for treating cancer, said methodcomprising administering to a subject in need thereof the compound ofFormula (IV) or a pharmaceutically acceptable salt thereof, in an amountsufficient to treat said cancer.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (IV) or a pharmaceutically acceptable salt thereof, in        an amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound of Formula (IV) or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

5.2.6 The Compounds of Formula (IVa)

As stated above, the present invention encompasses compounds having theFormula (IVa):

and pharmaceutically acceptable salts thereof,

wherein

-   -   R^(6a) is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more of the following groups: -halo,        —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′,        NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or        unsubstituted —C₁-C₆ alkyl;    -   m is an integer ranging from 0-10; and    -   n is an integer ranging from 2-10.

An illustrative example of a Compound of Formula (IVa) is the compoundhaving the formula:

Compound No. R^(6a) m n 17 Adamantyl 0 5or a pharmaceutically acceptable salt thereof.

A method for increasing the sensitivity of a cancer cell to thecytotoxic effects of radiotherapy, said method comprising contactingsaid cell with the compound of Formula (IVa) or pharmaceutically saltthereof, effective to increase the sensitivity of said cell to thecytotoxic effects of radiotherapy.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (IVa) or a pharmaceutically acceptable salt thereof, in        an amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound of Formula (IVa) or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

5.2.7 The Compounds of Formula (V)

As stated above, the present invention encompasses compounds having theFormula (V):

and pharmaceutically acceptable salts thereof,

wherein

-   -   Y is —C(O)CH₂SH or —NHC(O)CH₂SH;    -   R⁷ is —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl or -3- to        10-membered heterocycle, any of which may be unsubstituted or        substituted with one or more of the following groups: -halo,        —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′,        NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or        unsubstituted —C₁-C₆ alkyl; with the proviso that when n is 2,        R⁷ cannot be —C₃-C₇ cycloalkyl or -3- to 10-membered        heterocycle;    -   m is an integer ranging from 0-10; and    -   n is an integer ranging from 1-10.

The present invention also provides pharmaceutical compositionscomprising the compound of Formula (V) or a pharmaceutically acceptablesalt thereof, and a pharmaceutically acceptable carrier or vehicle.

A method for increasing the sensitivity of a cancer cell to thecytotoxic effects of radiotherapy, said method comprising contactingsaid cell with the compound of Formula (V) or pharmaceutically saltthereof, effective to increase the sensitivity of said cell to thecytotoxic effects of radiotherapy.

The invention also provides a method for treating cancer, said methodcomprising administering to a subject in need thereof the compound ofFormula (V) or a pharmaceutically acceptable salt thereof, in an amountsufficient to treat said cancer.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (V) or a pharmaceutically acceptable salt thereof, in an        amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound of Formula (V) or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

5.2.8 The Compounds of Formula (VI)

As stated above, the present invention encompasses compounds having theFormula (VI):

and pharmaceutically acceptable salts thereof,

wherein

-   -   each Z is independently —C(O)NHOH, —C(O)CH₂SH or —NHC(O)CH₂SH,        with the proviso that when both Z groups are —C(O)NHOH, the        phenyl group of said compound of formula (VI) is either ortho or        meta substituted;    -   m is an integer ranging from 1-10; and    -   n is an integer ranging from 1-10.

A subclass of the compounds of Formula (VI) is that wherein eachoccurrence of Z is —C(O)NHOH.

An illustrative Compound of Formula (VI) is the compound shown below:

Compound No. Z m n 18 —C(O)NHOH 6 6or a pharmaceutically acceptable salt thereof.

The present invention also provides pharmaceutical compositionscomprising the compound of Formula (VI) or a pharmaceutically acceptablesalt thereof, and a pharmaceutically acceptable carrier or vehicle.

A method for increasing the sensitivity of a cancer cell to thecytotoxic effects of radiotherapy, said method comprising contactingsaid cell with the compound of Formula (VI) or pharmaceutically saltthereof, effective to increase the sensitivity of said cell to thecytotoxic effects of radiotherapy.

The invention also provides a method for treating cancer, said methodcomprising administering to a subject in need thereof the compound ofFormula (VI) or a pharmaceutically acceptable salt thereof, in an amountsufficient to treat said cancer.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (VI) or a pharmaceutically acceptable salt thereof, in        an amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound of Formula (VI) or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

5.2.9 The Compounds of Formula (VII)

As stated above, the present invention encompasses compounds having theFormula (VII):

and pharmaceutically acceptable salts thereof,

wherein

-   -   each Y is independently —C(O)CH₂SH or —NHC(O)CH₂SH;    -   m is an integer ranging from 1-10; and    -   n is an integer ranging from 1-10.

The present invention also provides a method for increasing thesensitivity of a cancer cell to the cytotoxic effects of radiotherapy,said method comprising contacting said cell with the compound of Formula(VII) or pharmaceutically salt thereof, effective to increase thesensitivity of said cell to the cytotoxic effects of radiotherapy.

The invention also provides a method for treating cancer, said methodcomprising administering to a subject in need thereof the compound ofFormula (VII) or a pharmaceutically acceptable salt thereof, in anamount sufficient to treat said cancer.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (VII) or a pharmaceutically acceptable salt thereof, in        an amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound of Formula (VII) or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

5.2.10 The Compounds of Formula (VIII)

As stated above, the present invention encompasses compounds having theFormula (VIII):

and pharmaceutically acceptable salts thereof,

wherein

-   -   each R⁸ is independently —C₁-C₆ alkyl, aryl, —C₃-C₇ cycloalkyl        or -3- to 10-membered heterocycle, any of which may be        unsubstituted or substituted with one or more of the following        groups: -halo, —C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′,        —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′        is —H or unsubstituted —C₁-C₆ alkyl;    -   each G is independently —NH— or —CH₂—;    -   each J is independently —NH— or —CH₂—;    -   each m is independently an integer ranging from 1-10; and    -   each n is independently an integer ranging from 1-10.

Illustrative examples of Compounds of Formula (VIII) include thecompounds listed below:

Compound No. R⁸ G J m n 19 Phenyl —NH— —NH— 0 6 20 4-N(CH₃)₂-Phenyl —NH——NH— 1 6and pharmaceutically acceptable salts thereof.

The present invention also provides pharmaceutical compositionscomprising the compound of Formula (VIII) or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier orvehicle.

A method for increasing the sensitivity of a cancer cell to thecytotoxic effects of radiotherapy, said method comprising contactingsaid cell with the compound of Formula (VIII) or pharmaceutically saltthereof, effective to increase the sensitivity of said cell to thecytotoxic effects of radiotherapy.

The invention also provides a method for treating cancer, said methodcomprising administering to a subject in need thereof the compound ofFormula (VIII) or a pharmaceutically acceptable salt thereof, in anamount sufficient to treat said cancer.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (VIII) or a pharmaceutically acceptable salt thereof, in        an amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound of Formula (VIII) or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

5.2.11 The Compounds of Formula (IX)

As stated above, the present invention encompasses compounds having theFormula (IX)

and pharmaceutically acceptable salts thereof,

wherein

-   -   R⁹ is phenyl, which can be unsubstituted or substituted with one        or more of the following groups: -halo, —C₁-C₆ alkyl, —O—(C₁-C₆        alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′, N(R′)₂, —NHC(O)R′ or        —C(O)NHR′ groups wherein R′ is —H or unsubstituted —C₁-C₆ alkyl;        and    -   m is an integer ranging from 2-10.

A first subclass of the compounds of Formula (IX) is that wherein m is5.

A second subclass of the compounds of Formula (IX) is that wherein m is6.

A third subclass of the compounds of Formula (DC) is that wherein R⁹ is-phenyl.

A fourth subclass of the compounds of Formula (IX) is that wherein R⁹ is-4-N(CH₃)₂-phenyl.

A fifth subclass of the compounds of Formula (IX) is that wherein R⁹ is-4-biphenyl.

Illustrative examples of Compounds of Formula (IX) include the compoundslisted below:

Compound No. R⁹ 21 -phenyl 22 -4-N(CH₃)₂-phenyl 23 -4-biphenyl 24-4-N(CH₃)₂-phenyl 25 -phenyland pharmaceutically acceptable salts thereof.

The present invention also provides pharmaceutical compositionscomprising the compound of Formula (IX) or a pharmaceutically acceptablesalt thereof, and a pharmaceutically acceptable carrier or vehicle.

A method for increasing the sensitivity of a cancer cell to thecytotoxic effects of radiotherapy, said method comprising contactingsaid cell with the compound of Formula (IX) or pharmaceutically saltthereof, in an amount sufficient to increase the sensitivity of saidcell to the cytotoxic effects of radiotherapy.

The invention also provides a method for treating cancer, said methodcomprising administering to a subject in need thereof the compound ofFormula (IX) or a pharmaceutically acceptable salt thereof, in an amountsufficient to treat said cancer.

The invention also provides a method for treating cancer, said methodcomprising the steps of:

-   -   (a) administering to a subject in need thereof the compound of        Formula (IX) or a pharmaceutically acceptable salt thereof, in        an amount sufficient to sensitize a cancer cell to the cytotoxic        effects of radiotherapy; and    -   (b) administering to said subject an amount of radiotherapy        sufficient to treat said cancer.

The invention also provides a method for treating a neurologicaldisease, said method comprising administering to a subject in needthereof the compound of Formula (IX) or a pharmaceutically acceptablesalt thereof, in an amount sufficient to treat said neurologicaldisease.

For ease of reference, the compounds of Formulas (I), (Ia), (II), (III),(IV), (IVa), (V), (VI), (VII), (VIII) and (IX) will simply be referredto herein as the “Compounds of the Invention.”

5.3 Preparation of the Compounds of the Invention

The Compounds of the Invention may be prepared via the syntheticprocedure outlined below in Schemes 1-9. It will be apparent to oneskilled in the art how to prepare the scope of the Compounds of theInvention by choice of proper and relevant starting materials, syntheticintermediates and reagents.

Accordingly, Scheme 1 illustrates a method useful for making thecompounds of Formula (I).

The tosylate salt of an amine of formula 26 is treated with triphosgeneto provide an intermediate isocyanate which is reacted in situ with anamine of formula 27 to provide the urea of formula 28. The benzylprotecting group of compound 28 is removed using catalytic hydrogenationand the unmasked carboxylic acid is subsequently coupled with a benzylprotected hydroxylamine to provide the benzyl protected hydroxyamide offormula 29. Compound 29 is then debenzylated using catalytichydrogenation to provide the compound of Formula (I).

Scheme 2 shows a method useful for making the compounds of Formula (II)where Y is —NHC(O)CH₂SH.

The thiol group of methylthioglycolate 30 is protected as it's tritylderivative 31, which is subsequently couples with an alkyldiamine offormula 32 to provide amine intermediate 33. Intermediate 33 is thencoupled with an isocyanate of formula 34, and the trityl protectinggroup is removed to provide the Compound of Formula (II), where Y is—NHC(O)CH₂SH.

Scheme 3 shows a method useful for making the compounds of Formula (II)where Y is —C(O)CH₂SH.

Silylated acetic acid derivative 35 is converted to TBS protectedthioglycolic acid 36, which is treated with N,O-dimethylhydroxylaminehydrochloride to provide the N-methoxy-N-methyl amide 37. Compound 37 iscoupled with an alkanol bis-Grignard reagent of formula 38 to providealcohol 39, which is transformed to amine 40 using a variant of theGabriel amine synthesis. Amine 40 is then coupled with an isocyanate offormula 34, and the coupled product is subjected to an acid-catalyzeddeprotection of the silyl-protected thiol group to provide the compoundof formula (II), where Y is —C(O)CH₂SH.

Scheme 4 depicts methodology useful for preparing the compounds offormula (III).

The tosylate salt of an amine of formula 26 is treated with triphosgeneto provide an intermediate isocyanate which is reacted in situ with anamine of formula 41 to provide the urea of formula 42. The benzylprotecting group of compound 42 is removed using catalytic hydrogenationand the unmasked carboxylic acid is subsequently coupled with a benzylprotected hydroxylamine to provide the benzyl protected hydroxyamide offormula 43. Compound 43 is subsequently debenzylated using catalytichydrogenation to provide the compound of Formula (III).

Scheme 5 shows a method useful for making the compounds of Formula (IV).

An amine of general formula 44 is subjected to an acid-catalyzedcoupling with a cyclic anhydride of formula 45 in alcoholic solvent toprovide ester intermediate 46, which is then converted to thehydroxyamide of Formula (IV) via treatment with hydroxylaminehydrochloride in the presence of base.

Scheme 6 illustrates methodology useful for preparing the compounds offormula (V) where Y is —NHC(O)CH₂SH.

The thiol group of thioglycolic acid 47 is protected as it's tritylderivative 48, which is subsequently treated withN,O-dimethylhydroxylamine hydrochloride to provide theN-methoxy-N-methyl amide 49. Compound 49 is coupled with an alkanolbis-Grignard reagent of formula 50 to provide alcohol 51, which isoxidized to provide carboxylic acid 52. Compound 52 is coupled with anamine of formula 53 using EDCI and the thiol protecting group is removedusing TFA to provide the compound of Formula (V) where Y is—NHC(O)CH₂SH.

Scheme 7 illustrates methodology useful for preparing the compounds offormula (V) where Y is —C(O)CH₂SH.

The thiol group of thioglycolic acid 47 is protected as it's tritylderivative 48, which is subsequently coupled with an alkylamine offormula 54, followed by basic hydrolysis to yield the carboxylic acidintermediate of formula 55. Intermediate 55 is then coupled with analkylamine of formula 53, followed by removal of the trityl group toprovide the Compound of Formula (V), where Y is —NHC(O)CH₂SH.

Scheme 8 shows a method useful for preparing the compounds of formulas(VI) and (VII) wherein the integers m and n are the same.

A phenyldiamine of general formula 56 is coupled with an excess of acyclic anhydride of formula 57 to provide a diester intermediate offormula 58, which is subsequently converted to the dihydroxyamides ofFormulas (VI) and (VII) upon treatment with hydroxylamine hydrochloride.

Alternatively, Scheme 9 illustrates how Scheme 8 can be modified toprovide compound of Formulas (VI) and (VII) having different values of mand n by reacting compound 56 with one equivalent of the cyclicanhydride of formula 57 and reacting the product of this reaction withone equivalent of the cyclic anhydride of formula 59 to provide thediester intermediate of formula 60, which can be brought forward to thecompounds of formula (VI) and (VII) using the methodology shown inScheme 8.

It will be apparent to a person of ordinary skill in the art of organicsynthesis how to prepare the compounds of formulas (VI) and (VII) havingZ groups which are not identical by sequentially subjecting diamine 56to the any two of the chemical methodologies described in schemes 6, 7or 8 in proper stoiciometric amounts.

Scheme 10 illustrates methodology useful for preparing the compounds offormula (VIII).

The thiol groups of Compounds of Formulas (II) and (V) may beoxidatively self-coupled in the presence of triethylamine to provide thedisulfide compounds of Formula (VIII).

It will be apparent to a person of ordinary skill in the art of organicsynthesis how to prepare the compounds of formulas (VIII) andnon-identical R⁸ and/or G and/or J and/or m and/or n groups by using themethodology described in Scheme 10 to heterocouple two non-identicalcompounds of formula (II), two non-identical compounds of formula (V) ora compound of formula (II) and a compound of formula (V).

5.6 Pharmaceutical Compositions and Therapeutic Administration

In other aspects, the present invention provides a pharmaceuticalcomposition comprising an effective amount of a Compound of theInvention and a pharmaceutically acceptable carrier or vehicle. Thepharmaceutical compositions are suitable for veterinary or humanadministration.

The pharmaceutical compositions of the present invention can be in anyform that allows for the composition to be administered to a subject,said subject preferably being an animal, including, but not limited to ahuman, mammal, or non-human animal, such as a cow, horse, sheep, pig,fowl, cat, dog, mouse, rat, rabbit, guinea pig, etc., and is morepreferably a mammal, and most preferably a human.

The compositions of the invention can be in the form of a solid, liquidor gas (aerosol). Typical routes of administration may include, withoutlimitation, oral, topical, parenteral, sublingual, rectal, vaginal,ocular, and intranasal. Parenteral administration includes subcutaneousinjections, intravenous, intramuscular, intraperitoneal, intrapleural,intrasternal injection or infusion techniques. Preferably, thecompositions are administered parenterally, most preferablyintravenously. Pharmaceutical compositions of the invention can beformulated so as to allow a Compound of the Invention to be bioavailableupon administration of the composition to a subject. Compositions cantake the form of one or more dosage units, where for example, a tabletcan be a single dosage unit, and a container of a Compound of theInvention in aerosol form can hold a plurality of dosage units.

Materials used in preparing the pharmaceutical compositions can benon-toxic in the amounts used. It will be evident to those of ordinaryskill in the art that the optimal dosage of the active ingredient(s) inthe pharmaceutical composition will depend on a variety of factors.Relevant factors include, without limitation, the type of subject (e.g.,human), the overall health of the subject, the type of cancer thesubject is in need of treatment of, the use of the composition as partof a multi-drug regimen, the particular form of the Compound of theInvention, the manner of administration, and the composition employed.

The pharmaceutically acceptable carrier or vehicle may be particulate,so that the compositions are, for example, in tablet or powder form. Thecarrier(s) can be liquid, with the compositions being, for example, anoral syrup or injectable liquid. In addition, the carrier(s) can begaseous, so as to provide an aerosol composition useful in, e.g.,inhalatory administration.

The composition may be intended for oral administration, and if so, thecomposition is preferably in solid or liquid form, where semi-solid,semi-liquid, suspension and gel forms are included within the formsconsidered herein as either solid or liquid.

As a solid composition for oral administration, the composition can beformulated into a powder, granule, compressed tablet, pill, capsule,chewing gum, wafer or the like form. Such a solid composition typicallycontains one or more inert diluents. In addition, one or more of thefollowing can be present: binders such as ethyl cellulose,carboxymethylcellulose, microcrystalline cellulose, or gelatin;excipients such as starch, lactose or dextrins, disintegrating agentssuch as alginic acid, sodium alginate, Primogel, corn starch and thelike; lubricants such as magnesium stearate or Sterotex; glidants suchas colloidal silicon dioxide; sweetening agents such as sucrose orsaccharin, a flavoring agent such as peppermint, methyl salicylate ororange flavoring, and a coloring agent.

When the pharmaceutical composition is in the form of a capsule, e.g., agelatin capsule, it can contain, in addition to materials of the abovetype, a liquid carrier such as polyethylene glycol, cyclodextrin or afatty oil.

The pharmaceutical composition can be in the form of a liquid, e.g., anelixir, syrup, solution, emulsion or suspension. The liquid can beuseful for oral administration or for delivery by injection. Whenintended for oral administration, a composition can comprise one or moreof a sweetening agent, preservatives, dye/colorant and flavor enhancer.In a composition for administration by injection, one or more of asurfactant, preservative, wetting agent, dispersing agent, suspendingagent, buffer, stabilizer and isotonic agent can also be included.

The liquid compositions of the invention, whether they are solutions,suspensions or other like form, can also include one or more of thefollowing: sterile diluents such as water for injection, salinesolution, preferably physiological saline, Ringer's solution, isotonicsodium chloride, fixed oils such as synthetic mono or digylcerides whichcan serve as the solvent or suspending medium, polyethylene glycols,glycerin, cyclodextrin, propylene glycol or other solvents;antibacterial agents such as benzyl alcohol or methyl paraben;antioxidants such as ascorbic acid or sodium bisulfite; chelating agentssuch as ethylenediaminetetraacetic acid; buffers such as acetates,citrates or phosphates and agents for the adjustment of tonicity such assodium chloride or dextrose. A parenteral composition can be enclosed inampoule, a disposable syringe or a multiple-dose vial made of glass,plastic or other material. Physiological saline is a preferred adjuvant.An injectable composition is preferably sterile.

The amount of the Compound of the Invention that is effective in thetreatment of a particular disorder or condition will depend on thenature of the disorder or condition, and can be determined by standardclinical techniques. In addition, in vitro or in vivo assays canoptionally be employed to help identify optimal dosage ranges. Theprecise dose to be employed in the compositions will also depend on theroute of administration, and the seriousness of the disease or disorder,and should be decided according to the judgment of the practitioner andeach patient's circumstances.

The pharmaceutical compositions comprise an effective amount of aCompound of the Invention such that a suitable dosage will be obtained.Typically, this amount is at least 0.01% of a Compound of the Inventionby weight of the composition. When intended for oral administration,this amount can be varied to be between 0.1% and 80% by weight of thecomposition. Preferred oral compositions can comprise from between 4%and 50% of the Compound of the Invention by weight of the composition.Preferred compositions of the present invention are prepared so that aparenteral dosage unit contains from between 0.01% and 2% by weight ofthe Compound of the Invention.

Generally, the dosage of a Compound of the Invention administered to asubject is typically between 0.1 mg/kg and 100 mg/kg of the subject'sbody weight. In one embodiment, the dosage administered to a subject isbetween 0.5 mg/kg and 50 mg/kg of the subject's body weight, morepreferably between 1 mg/kg and 25 mg/kg of the subject's body weight.

In a specific embodiment, when the Compounds of the Invention are usedin combination with radiotherapy, a Compound of the Invention can beadministered in amounts that result in concentrations in the fluid of atarget tissue that are less than about twice the IC₅₀ concentration forthe particular compound, more preferably about equal to the IC₅₀concentration. The IC₅₀ concentration is defined as the concentration ofthe Compound of the Invention that kills 50% of cells followingtreatment with the Compound of the Invention.

In another embodiment, the Compounds of the Invention may beadministered at amounts lower than the IC₅₀ concentration, such as about50% of the IC₅₀ concentration, about 40% of the IC₅₀ concentration,about 30% of the IC₅₀ concentration, about 20% of the IC₅₀concentration, about 10% or about 5% of the IC₅₀ concentration, at thetarget tissue.

In still another embodiment, the Compounds of the Invention may beadministered locally so that the concentration at the target tissue isin the effective range and the concentration in non-target tissue isminimized.

In another embodiment, the dosage of the Compound of the Inventionresults in a concentration at a target tissue that does not promoteapoptosis of cells in culture yet is effective in increasing cell deathin neoplastic cells exposed to radiation or recognized chemotherapeuticchemical agents. Concentrations that produce these effects can bedetermined for a Compound of the Invention by one of skill in the artusing markers of apoptosis, including, but not limited to, the apoptoticindex and caspase activities.

The Compounds of the Invention can be administered by any convenientroute, for example by infusion or bolus injection, by absorption throughepithelial or mucocutaneous linings (e.g., oral mucosa, rectal andintestinal mucosa, etc.). Administration can be systemic or local.Various delivery systems are known, e.g., microparticles, microcapsules,capsules, etc., and may be useful for administering a Compound of theInvention. In certain embodiments, more than one Compound of theInvention is administered to a subject. Methods of administration mayinclude, but are not limited to, oral administration and parenteraladministration; parenteral administration including, but not limited to,intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous;intranasal, epidural, sublingual, intranasal, intracerebral,intraventricular, intrathecal, intravaginal, transdermal, rectally, byinhalation, or topically to the ears, nose, eyes, or skin. The preferredmode of administration is left to the discretion of the practitioner,and will depend in-part upon the site of the medical condition (such asthe site of cancer, a cancerous tumor or a pre-cancerous condition).

In one embodiment, the Compounds of the Invention are administeredorally.

In another embodiment, the Compounds of the Invention are administeredparenterally.

In still another embodiment, the Compounds of the Invention areadministered intravenously.

In specific embodiments, it can be desirable to administer one or moreCompounds of the Invention locally to the area in need of treatment.This can be achieved, for example, and not by way of limitation, bylocal infusion during surgery; topical application, e.g., in conjunctionwith a wound dressing after surgery; by injection; by means of acatheter; by means of a suppository; or by means of an implant, theimplant being of a porous, non-porous, or gelatinous material, includingmembranes, such as sialastic membranes, or fibers. In one embodiment,administration can be by direct injection at the site (or former site)of a cancer, tumor, or precancerous tissue. In certain embodiments, itcan be desirable to introduce one or more Compounds of the Inventioninto the central nervous system by any suitable route, includingintraventricular and intrathecal injection. Intraventricular injectioncan be facilitated by an intraventricular catheter, for example,attached to a reservoir, such as an Ommaya reservoir.

Pulmonary administration can also be employed, e.g., by use of aninhaler or nebulizer, and formulation with an aerosolizing agent, or viaperfusion in a fluorocarbon or synthetic pulmonary surfactant. Incertain embodiments, the Compounds of the Invention can be formulated asa suppository, with traditional binders and carriers such astriglycerides.

In one embodiment, the Compounds of the Invention can be delivered in avesicle, in particular a liposome (see Langer, Science 249:1527-1533(1990); Treat et al., in Liposomes in the Therapy of Infectious Diseaseand Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp.353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; see generallyibid.).

In yet another embodiment, the Compounds of the Invention can bedelivered in a controlled release system. In one embodiment, a pump canbe used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201(1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., N. Engl.J. Med. 321:574 (1989)). In another embodiment, polymeric materials canbe used (see Medical Applications of Controlled Release, Langer and Wise(eds.), CRC Pres., Boca Raton, Fla. (1974); Controlled DrugBioavailability, Drug Product Design and Performance, Smolen and Ball(eds.), Wiley, New York (1984); Ranger and Peppas, J. Macromol. Sci.Rev. Macromol. Chem. 23:61 (1983); see also Levy et al., Science 228:190(1985); During et al., Ann. Neurol. 25:351 (1989); Howard et al., J.Neurosurg. 71:105 (1989)). In yet another embodiment, acontrolled-release system can be placed in proximity of the target ofthe Compounds of the Invention, e.g., the brain, thus requiring only afraction of the systemic dose (see, e.g., Goodson, in MedicalApplications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)).Other controlled-release systems discussed in the review by Langer(Science 249:1527-1533 (1990)) can be used.

The term “carrier” refers to a diluent, adjuvant or excipient, withwhich a Compound of the Invention is administered. Such pharmaceuticalcarriers can be liquids, such as water and oils, including those ofpetroleum, animal, vegetable or synthetic origin, such as peanut oil,soybean oil, mineral oil, sesame oil and the like. The carriers can besaline, gum acacia, gelatin, starch paste, talc, keratin, colloidalsilica, urea, and the like. In addition, auxiliary, stabilizing,thickening, lubricating and coloring agents can be used. In oneembodiment, when administered to a subject, the Compounds of theInvention and pharmaceutically acceptable carriers are sterile. Water isa preferred carrier when the Compound of the Invention is administeredintravenously. Saline solutions and aqueous dextrose and glycerolsolutions can also be employed as liquid carriers, particularly forinjectable solutions. Suitable pharmaceutical carriers also includeexcipients such as starch, glucose, lactose, sucrose, gelatin, malt,rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate,talc, sodium chloride, dried skim milk, glycerol, propylene, glycol,water, ethanol and the like. The present compositions, if desired, canalso contain minor amounts of wetting or emulsifying agents, or pHbuffering agents.

The present compositions can take the form of solutions, suspensions,emulsion, tablets, pills, pellets, capsules, capsules containingliquids, powders, sustained-release formulations, suppositories,emulsions, aerosols, sprays, suspensions, or any other form suitable foruse. In one embodiment, the pharmaceutically acceptable carrier is acapsule (see e.g., U.S. Pat. No. 5,698,155). Other examples of suitablepharmaceutical carriers are described in “Remington's PharmaceuticalSciences” by E. W. Martin.

Sustained or directed release compositions that may be formulatedinclude, but are not limited to liposomes or other formulations whereinthe active component is protected with differentially degradablecoatings, e.g., by microencapsulation, multiple coatings, etc. It isalso possible to freeze-dry the new compositions and use thelyophilizates obtained, for example, for the preparation of products forinjection.

In a preferred embodiment, the Compounds of the Invention are formulatedin accordance with routine procedures as a pharmaceutical compositionadapted for intravenous administration to animals, particularly humanbeings. Typically, the carriers or vehicles for intravenousadministration are sterile isotonic aqueous buffer solutions. Wherenecessary, the compositions can also include a solubilizing agent.Compositions for intravenous administration can optionally comprise alocal anesthetic such as lignocaine to ease pain at the site of theinjection. Generally, the ingredients are supplied either separately ormixed together in unit dosage form, for example, as a dry lyophilizedpowder or water free concentrate in a hermetically sealed container suchas an ampoule or sachette indicating the quantity of active agent. Wherea Compound of the Invention is to be administered by infusion, it can bedispensed, for example, with an infusion bottle containing sterilepharmaceutical grade water or saline. Where the Compound of theInvention is administered by injection, an ampoule of sterile water forinjection or saline can be provided so that the ingredients can be mixedprior to administration.

Compositions for oral delivery can be in the form of tablets, lozenges,aqueous or oily suspensions, granules, powders, emulsions, capsules,syrups, or elixirs, for example. Orally administered compositions cancontain one or more optionally agents, for example, sweetening agentssuch as fructose, aspartame or saccharin; flavoring agents such aspeppermint, oil of wintergreen, or cherry; coloring agents; andpreserving agents, to provide a pharmaceutically palatable preparation.Moreover, where in tablet or pill form, the compositions can be coatedto delay disintegration and absorption in the gastrointestinal tractthereby providing a sustained action over an extended period of time.Selectively permeable membranes surrounding an osmotically activedriving complex are also suitable for orally administered compositionsof the invention. In these later platforms, fluid from the environmentsurrounding the capsule is imbibed by the driving complex, which swellsto displace the agent or agent composition through an aperture. Thesedelivery platforms can provide an essentially zero order deliveryprofile as opposed to the spiked profiles of immediate releaseformulations. A time-delay material such as glycerol monostearate orglycerol stearate can also be used. Oral compositions can includestandard carriers such as mannitol, lactose, starch, magnesium stearate,sodium saccharine, cellulose, magnesium carbonate, etc. Such carriersare preferably of pharmaceutical grade.

The pharmaceutical compositions of the invention can be intended fortopical administration, in which case the carrier can be in the form ofa solution, emulsion, ointment or gel base. The base, for example, cancomprise one or more of the following: petrolatum, lanolin, polyethyleneglycols, beeswax, mineral oil, diluents such as water and alcohol, andemulsifiers and stabilizers. Thickening agents can be present in acomposition for topical administration. If intended for transdermaladministration, the composition can be in the form of a transdermalpatch or an iontophoresis device. Topical formulations can comprise aconcentration of a Compound of the Invention of from between 0.01% and10% w/v (weight per unit volume of composition).

The compositions can include various materials that modify the physicalform of a solid or liquid dosage unit. For example, the composition caninclude materials that form a coating shell around the activeingredients. The materials that form the coating shell are typicallyinert, and can be selected from, for example, sugar, shellac, and otherenteric coating agents. Alternatively, the active ingredients can beencased in a gelatin capsule.

The compositions can consist of gaseous dosage units, e.g., it can be inthe form of an aerosol. The term aerosol is used to denote a variety ofsystems ranging from those of colloidal nature to systems consisting ofpressurized packages. Delivery can be by a liquefied or compressed gasor by a suitable pump system that dispenses the active ingredients.Aerosols of the compositions can be delivered in single phase,bi-phasic, or tri-phasic systems in order to deliver the composition.Delivery of the aerosol includes the necessary container, activators,valves, subcontainers, Spacers and the like, which together can form akit. Preferred aerosols can be determined by one skilled in the art,without undue experimentation.

Whether in solid, liquid or gaseous form, the compositions of thepresent invention can comprise an additional therapeutically activeagent selected from among those including, but not limited to, anadditional anticancer agent, an antiemetic agent, a hematopoietic colonystimulating factor, an anti-depressant and an analgesic agent.

The pharmaceutical compositions can be prepared using methodology wellknown in the pharmaceutical art. For example, a composition intended tobe administered by injection can be prepared by combining a Compound ofthe Invention with water so as to form a solution. A surfactant can beadded to facilitate the formation of a homogeneous solution orsuspension. Surfactants are complexes that can non-covalently interactwith a Compound of the Invention so as to facilitate dissolution orhomogeneous suspension of the Compound of the Invention in the aqueousdelivery system.

In one embodiment, the pharmaceutical compositions of the presentinvention may comprise one or more additional anticancer agents.

In another embodiment, the pharmaceutical compositions of the presentinvention can be administered prior to, at the same time as, or after anadditional anticancer agent, or on the same day, or within 1 hour, 2hours, 12 hours, 24 hours, 48 hours, 72 hours, 1 week, 2 weeks, 3 weeksor 4 weeks of each other.

In one embodiment, the pharmaceutical compositions of the presentinvention may comprise one or more known therapeutically active agents.

In another embodiment, the pharmaceutical compositions of the presentinvention can be administered prior to, at the same time as, or after anantiemetic agent, or on the same day, or within 1 hour, 2 hours, 12hours, 24 hours, 48 hours or 72 hours of each other.

In another embodiment, the pharmaceutical compositions of the presentinvention can be administered prior to, at the same time as, or after ahematopoietic colony stimulating factor, or on the same day, or within 1hour, 2 hours, 12 hours, 24 hours, 48 hours, 72 hours, 1 week, 2 weeks,3 weeks or 4 weeks of each other.

In another embodiment, the pharmaceutical compositions of the presentinvention can be administered prior to, at the same time as, or after anopioid or non-opioid analgesic agent, or on the same day, or within 1hour, 2 hours, 12 hours, 24 hours, 48 hours or 72 hours of each other.

In another embodiment, the pharmaceutical compositions of the presentinvention can be administered prior to, at the same time as, or after ananti-depressant agent, or on the same day, or within 1 hour, 2 hours, 12hours, 24 hours, 48 hours or 72 hours of each other.

5.7 Kits

The invention encompasses kits that can simplify the administration ofthe Compounds of the Invention or composition of the invention to asubject.

A typical kit of the invention comprises unit dosages of the Compoundsof the Invention. In one embodiment, the unit dosage form is in acontainer, which can be sterile, containing an effective amount of oneof the Compounds of the Invention and a pharmaceutically acceptablecarrier or vehicle. In another embodiment, the unit dosage form is in acontainer containing an effective amount of one of the Compounds of theInvention as a lyophilate. In this instance, the kit can furthercomprise another container which contains a solution useful for thereconstitution of the lyophilate. The kit can also comprise a label orprinted instructions for use of the Compounds of the Invention. In oneembodiment, the kit comprises multiple containers: (a) a first containercontaining an unit dosage form of Compound of the Invention, and (b) oneor more additional containers each containing a unit dosage form of oneor more additional anticancer agents or pharmaceutically acceptablesalts thereof. In another embodiment the kit comprises a containercontaining a therapeutically active agent such as an antiemetic agent, ahematopoietic colony-stimulating factor, an analgesic agent or ananxiolytic agent.

In a further embodiment, the kit comprises a unit dosage form of apharmaceutical composition of the invention.

Kits of the invention can further comprise one or more devices that areuseful for administering the unit dosage forms of the Compounds of theInvention or a pharmaceutical composition of the invention. Examples ofsuch devices include, but are not limited to, a syringe, a drip bag, apatch or an enema, which optionally contain the unit dosage forms.

5.8 Therapeutic Uses

The Compounds of the Invention are useful for treating cancer andneurological diseases. The Compounds of the Invention are also usefulfor increasing the sensitivity of a cancer cell to the cytotoxic effectsof radiotherapy.

5.8.1 Treatment of Cancer

Cancer can be treated or prevented by administration of amounts of theCompounds of the invention that are effective to treat cancer or byadministration of a pharmaceutical composition comprising amounts of theCompounds of the invention that are effective to treat cancer.

5.8.1.1 Therapeutic Methods

In a preferred embodiment, the present invention provides methods fortreating cancer, including but not limited to: killing a cancer cell orneoplastic cell; inhibiting the growth of a cancer cell or neoplasticcell; inhibiting the replication of a cancer cell or neoplastic cell; orameliorating a symptom thereof, said methods comprising administering toa subject in need thereof an amount of the Compounds of the inventioneffective to treat cancer.

In one embodiment, the invention provides a method for treating cancer,said method comprising administering to a subject in need thereof anamount of a Compound of the Invention or a pharmaceutically acceptablesalt thereof, said amount sufficient to treat said cancer.

In another embodiment, the invention provides a method for increasingthe sensitivity of a cancer cell to the cytotoxic effects ofradiotherapy, said method comprising contacting said cell with aCompound of the Invention or a pharmaceutically acceptable salt thereof,in an amount sufficient to increase the sensitivity of said cell to thecytotoxic effects of radiotherapy.

In a further embodiment, the present invention provides a method fortreating cancer, said method comprising: (a) administering to a subjectin need thereof an amount of a Compound of the Invention; and (b)administering to said subject an amount of radiotherapy. In oneembodiment, the amounts administered are each effective to treat cancer.In another specific embodiment, the amounts are together effective totreat cancer. The Compound of the Invention and radiotherapy can actadditively or synergistically.

In another embodiment, the invention provides a method for treatingcancer, said method comprising administering to a subject in needthereof a pharmaceutical composition comprising an amount of a Compoundof the Invention effective to treat cancer.

The combination therapy of the invention can be used accordingly in avariety of settings for the treatment of various cancers.

In a specific embodiment, the subject in need of treatment haspreviously undergone treatment for cancer. Such previous treatmentsinclude, but are not limited to, prior chemotherapy, radiotherapy,surgery, or immunotherapy, such as cancer vaccines.

In another embodiment, the cancer being treated is a cancer which hasdemonstrated sensitivity to radiotherapy or is known to be responsive toradiotherapy. Such cancers include, but are not limited to,Non-Hodgkin's lymphoma, Hodgkin's disease, Ewing's sarcoma, testicularcancer, prostate cancer, ovarian cancer, bladder cancer, larynx cancer,cervical cancer, nasopharynx cancer, breast cancer, colon cancer,pancreatic cancer, head and neck cancer, esophogeal cancer, rectalcancer, small-cell lung cancer, non-small cell lung cancer, braintumors, or other CNS neoplasms.

In still another embodiment, the cancer being treated is a cancer whichhas demonstrated resistance to radiotherapy or is known to be refractoryto radiotherapy. A cancer may be determined to be refractory to atherapy when at least some significant portion of the cancer cells arenot killed or their cell division are not arrested in response totherapy. Such a determination can be made either in vivo or in vitro byany method known in the art for assaying the effectiveness of treatmenton cancer cells, using the art-accepted meanings of “refractory” in sucha context. In a specific embodiment, a cancer is refractory where thenumber of cancer cells has not been significantly reduced, or hasincreased.

Other cancers that can be treated with the Compounds and methods of theInvention include, but are not limited to, cancers disclosed below inTable 1 and metastases thereof

TABLE 1 Solid tumors, including but not limited to: fibrosarcomamyxosarcoma liposarcoma chondrosarcoma osteogenic sarcoma chordomaangiosarcoma endotheliosarcoma lymphangiosarcomalymphangioendotheliosarcoma synovioma mesothelioma Ewing's tumorleiomyosarcoma rhabdomyosarcoma colon cancer colorectal cancer kidneycancer pancreatic cancer bone cancer breast cancer ovarian cancerprostate cancer esophageal cancer stomach cancer oral cancer nasalcancer throat cancer squamous cell carcinoma basal cell carcinomaadenocarcinoma sweat gland carcinoma sebaceous gland carcinoma papillarycarcinoma papillary adenocarcinomas cystadenocarcinoma medullarycarcinoma bronchogenic carcinoma renal cell carcinoma hepatoma bile ductcarcinoma choriocarcinoma seminoma embryonal carcinoma Wilms' tumorcervical cancer uterine cancer testicular cancer small cell lungcarcinoma bladder carcinoma lung cancer epithelial carcinoma gliomaglioblastoma multiforme astrocytoma medulloblastoma craniopharyngiomaependymoma pinealoma hemangioblastoma acoustic neuroma oligodendrogliomameningioma skin cancer melanoma neuroblastoma retinoblastoma blood-bornecancers, including but not limited to: acute lymphoblastic leukemia(“ALL”) acute lymphoblastic B-cell leukemia acute lymphoblastic T-cellleukemia acute myeloblastic leukemia (“AML”) acute promyelocyticleukemia (“APL”) acute monoblastic leukemia acute erythroleukemicleukemia acute megakaryoblastic leukemia acute myelomonocytic leukemiaacute nonlymphocyctic leukemia acute undifferentiated leukemia chronicmyelocytic leukemia (“CML”) chronic lymphocytic leukemia (“CLL”) hairycell leukemia multiple myeloma acute and chronic leukemias:lymphoblastic myelogenous lymphocytic myelocytic leukemias Lymphomas:Hodgkin's disease non-Hodgkin's Lymphoma Multiple myeloma Waldenström'smacroglobulinemia Heavy chain disease Polycythemia vera

In one embodiment, the cancer is selected from the group consisting ofNon-Hodgkin's lymphoma, Hodgkin's disease, Ewing's sarcoma, testicularcancer, prostate cancer, ovarian cancer, bladder cancer, larynx cancer,cervical cancer, nasopharynx cancer, breast cancer, colon cancer,pancreatic cancer, head and neck cancer, esophogeal cancer, rectalcancer, small-cell lung cancer, non-small cell lung cancer, braintumors, and other CNS neoplasms.

5.8.1.2 Prophylactic Methods

The Compounds of the Invention can also be administered to preventprogression to a neoplastic or malignant state, including but notlimited to the cancers listed in Table 1. Such prophylactic use isindicated in conditions known or suspected of preceding progression toneoplasia or cancer, in particular, where non-neoplastic cell growthconsisting of hyperplasia, metaplasia, or most particularly, dysplasiahas occurred (for review of such abnormal growth conditions, see Robbinsand Angell, 1976, Basic Pathology, 2d Ed., W.B. Saunders Co.,Philadelphia, pp. 68-79). Hyperplasia is a form of controlled cellproliferation involving an increase in cell number in a tissue or organ,without significant alteration in structure or function. For example,endometrial hyperplasia often precedes endometrial cancer andprecancerous colon polyps often transform into cancerous lesions.Metaplasia is a form of controlled cell growth in which one type ofadult or fully differentiated cell substitutes for another type of adultcell. Metaplasia can occur in epithelial or connective tissue cells. Atypical metaplasia involves a somewhat disorderly metaplasticepithelium. Dysplasia is frequently a forerunner of cancer, and is foundmainly in the epithelia; it is the most disorderly form ofnon-neoplastic cell growth, involving a loss in individual celluniformity and in the architectural orientation of cells. Dysplasticcells often have abnormally large, deeply stained nuclei, and exhibitpleomorphism. Dysplasia characteristically occurs where there existschronic irritation or inflammation, and is often found in the cervix,respiratory passages, oral cavity, and gall bladder.

Alternatively or in addition to the presence of abnormal cell growthcharacterized as hyperplasia, metaplasia, or dysplasia, the presence ofone or more characteristics of a transformed phenotype, or of amalignant phenotype, displayed in vivo or displayed in vitro by a cellsample from a subject, can indicate the desirability ofprophylactic/therapeutic administration of the composition of theinvention. Such characteristics of a transformed phenotype includemorphology changes, looser substratum attachment, loss of contactinhibition, loss of anchorage dependence, protease release, increasedsugar transport, decreased serum requirement, expression of fetalantigens, disappearance of the 250,000 dalton cell surface protein, etc.(see also id., at pp. 84-90 for characteristics associated with atransformed or malignant phenotype).

In a specific embodiment, leukoplakia, a benign-appearing hyperplasticor dysplastic lesion of the epithelium, or Bowen's disease, a carcinomain situ, are pre-neoplastic lesions indicative of the desirability ofprophylactic intervention.

In another embodiment, fibrocystic disease (cystic hyperplasia, mammarydysplasia, particularly adenosis (benign epithelial hyperplasia)) isindicative of the desirability of prophylactic intervention.

The prophylactic use of the compounds and methods of the presentinvention are also indicated in some viral infections that may lead tocancer. For example, human papilloma virus can lead to cervical cancer(see, e.g., Hernandez-Avila et al., Archives of Medical Research (1997)28:265-271), Epstein-Barr virus (EBV) can lead to lymphoma (see, e.g.,Herrmann et al., J Pathol (2003) 199(2):140-5), hepatitis B or C viruscan lead to liver carcinoma (see, e.g., El-Serag, J Clin Gastroenterol(2002) 35(5 Suppl 2):S72-8), human T cell leukemia virus (HTLV)-I canlead to T-cell leukemia (see e.g., Mortreux et al., Leukemia (2003)17(1):26-38), human herpesvirus-8 infection can lead to Kaposi's sarcoma(see, e.g., Kadow et al., Curr Opin Investig Drugs (2002) 3(11):1574-9),and Human Immune deficiency Virus (HIV) infection contribute to cancerdevelopment as a consequence of immunodeficiency (see, e.g., Dal Maso etal., Lancet Oncol (2003) 4(2):110-9).

In other embodiments, a subject which exhibits one or more of thefollowing predisposing factors for malignancy can treated byadministration of the compounds or methods of the invention: achromosomal translocation associated with a malignancy (e.g., thePhiladelphia chromosome for chronic myelogenous leukemia, t(14; 18) forfollicular lymphoma, etc.), familial polyposis or Gardner's syndrome(possible forerunners of colon cancer), benign monoclonal gammopathy (apossible forerunner of multiple myeloma), a first degree kinship withpersons having a cancer or precancerous disease showing a Mendelian(genetic) inheritance pattern (e.g., familial polyposis of the colon,Gardner's syndrome, hereditary exostosis, polyendocrine adenomatosis,medullary thyroid carcinoma with amyloid production andpheochromocytoma, Peutz-Jeghers syndrome, neurofibromatosis of VonRecklinghausen, retinoblastoma, carotid body tumor, cutaneousmelanocarcinoma, intraocular melanocarcinoma, xeroderma pigmentosum,ataxia telangiectasia, Chediak-Higashi syndrome, albinism, Fanconi'saplastic anemia, and Bloom's syndrome; see Robbins and Angell, 1976,Basic Pathology, 2d Ed., W.B. Saunders Co., Philadelphia, pp. 112-113)etc.), and exposure to carcinogens (e.g., smoking, and inhalation of orcontacting with certain chemicals).

In another specific embodiment, the compounds and methods of theinvention are administered to a human subject to prevent progression tobreast, colon, ovarian, or cervical cancer.

5.8.1.3 Multi-Modality Therapy for Cancer

The Compounds of the Invention can be administered to a subject that hasundergone or is currently undergoing one or more additional anticancertreatment modalities including, but not limited to, chemotherapy,radiotherapy, surgery or immunotherapy, such as cancer vaccines.

In one embodiment, the invention provides methods for treating cancercomprising (a) administering to a subject in need thereof an amount of acombination therapy of the invention; and (b) administering to saidsubject one or more additional anticancer treatment modalitiesincluding, but not limited to, radiotherapy, chemotherapy, surgery orimmunotherapy, such as a cancer vaccine. In one embodiment, theadministering of step (a) is done prior to the administering of step(b). In another embodiment, the administering of step (a) is donesubsequent to the administering of step (b). In still anotherembodiment, the administering of step (a) is done concurrently with theadministering of step (b).

In one embodiment, the additional anticancer treatment modality ischemotherapy.

In another embodiment, the additional anticancer treatment modality issurgery.

In still another embodiment, the additional anticancer treatmentmodality is immunotherapy, such as cancer vaccines.

In one embodiment, the Compound of the Invention or a pharmaceuticallyacceptable salt thereof is administered adjunctively with the additionalanticancer treatment modality.

In another embodiment, the Compound of the Invention or apharmaceutically acceptable salt thereof acts synergistically withradiotherapy.

In a preferred embodiment, the additional anticancer treatment modalityis radiotherapy. In the methods of the present invention, anyradiotherapy protocol can be used depending upon the type of cancer tobe treated. For example, but not by way of limitation, X-ray radiationcan be administered; in particular, high-energy megavoltage (radiationof greater that 1 MeV energy) can be used for deep tumors, and electronbeam and orthovoltage X-ray radiation can be used for skin cancers.Gamma-ray emitting radioisotopes, such as radioactive isotopes ofradium, cobalt and other elements, can also be administered.Illustrative radiotherapy protocols useful in the present inventioninclude, but are not limited to, stereotactic methods where multiplesources of low dose radiation are simultaneously focused into a tissuevolume from multiple angles; “internal radiotherapy,” such asbrachytherapy, interstitial irradiation, and intracavitary irradiation,which involves the placement of radioactive implants directly in a tumoror other target tissue; intraoperative irradiation, in which a largedose of external radiation is directed at the target tissue which isexposed during surgery; and particle beam radiotherapy, which involvesthe use of fast-moving subatomic particles to treat localized cancers.

In a preferred embodiment, the Compound of the Invention or apharmaceutically acceptable salt thereof is administered prior to theadministration of radiotherapy.

In another preferred embodiment, the Compound of the Invention or apharmaceutically acceptable salt thereof is administered adjunctivelywith radiotherapy.

The Compound of the Invention and the additional treatment modalities ofthe combination therapies of the invention can act additively orsynergistically (i.e., the combination of an Compound of the Inventionor a pharmaceutically acceptable salt thereof, and an additionalanticancer treatment modality is more effective than their additiveeffects when each are administered alone). A synergistic combinationpermits the use of lower dosages of the Compound of the Invention and/orthe additional treatment modality and/or less frequent administration ofthe Compound of the Invention and/or additional treatment modality to asubject with cancer. The ability to utilize lower dosages of a Compoundof the Invention and/or an additional treatment modality and/or toadminister a Compound of the Invention and said additional treatmentmodality less frequently can reduce the toxicity associated with theadministration of a Compound of the Invention and/or the additionaltreatment modality to a subject without reducing the efficacy of aCompound of the Invention and/or the additional treatment modality inthe treatment of cancer. In addition, a synergistic effect can result inthe improved efficacy of the treatment of cancer and/or the reduction ofadverse or unwanted side effects associated with the administration of aCompound of the Invention and/or an additional anticancer treatmentmodality as monotherapy.

In one embodiment, the Compounds of the Invention may actsynergistically with radiotherapy when administered in doses typicallyemployed when such agents are used alone for the treatment of cancer. Inanother embodiment, the Compounds of the Invention may actsynergistically with radiotherapy when administered in doses that areless than doses typically employed when such agents are used asmonotherapy for the treatment of cancer.

In one embodiment, radiotherapy may act synergistically with a Compoundof the Invention when administered in doses typically employed whenradiotherapy is used as monotherapy for the treatment of cancer. Inanother embodiment, radiotherapy may act synergistically with a Compoundof the Invention when administered in doses that are less than dosestypically employed when radiotherapy is used as monotherapy for thetreatment of cancer.

In a specific embodiment, the Compounds of the Invention act as HDACinhibitors.

The effectiveness of the use of the Compounds of the Invention as HDACinhibitors for sensitizing cancer cells to the effect of radiotherapycan be determined by the in vitro and/or in vivo determination ofpost-treatment survival using techniques known in the art. In oneembodiment, for in vitro determinations, exponentially growing cells canbe exposed to known doses of radiation and the survival of the cellsmonitored. Irradiated cells are plated and cultured for about 14-about21 days, and the colonies are stained. The surviving fraction is thenumber of colonies divided by the plating efficiency of unirradiatedcells. Graphing the surviving fraction on a log scale versus theabsorbed dose on a linear scale generates a survival curve. Survivalcurves generally show an exponential decrease in the fraction ofsurviving cells at higher radiation doses after an initial shoulderregion in which the dose is sublethal. A similar protocol can be usedfor chemical agents when used in the combination therapies of theinvention.

Inherent radiosensitivity of tumor cells and environmental influences,such as hypoxia and host immunity, can be further assessed by in vivostudies. The growth delay assay is commonly used. This assay measuresthe time interval required for a tumor exposed to radiation to regrow toa specified volume. The dose required to control about 50% of tumors isdetermined by the TCD₅₀ assay.

In vivo assay systems typically use transplantable solid tumor systemsin experimental subjects. Radiation survival parameters for normaltissues as well as for tumors can be assayed using in vivo methods knownin the art.

Two mathematical models are commonly employed to analyze radiationsurvival data. A first model is the multi-target model. In thisanalysis, the reciprocal of the slope of the survival curve is definedas Do, the radiosensitivity of the cell population or tissue underinvestigation. Do is the dose required to reduce the surviving fractionto about 37% in the exponential portion of the survival curve. Theextrapolation of the linear portion of the curve to the y-intercept isdenoted n. The width of the shoulder region is represented by drawing aline from the 100% survival point to the extrapolation line, this widthis denoted Dq. Dq is the quasi-threshold dose, or the point at which thereduction in surviving fraction as a function of radiation dosagebecomes exponential. The Dq value can also provide an estimate of anadditional total dose required for each division of a single dosetherapy into fractional doses. The additional dose is required toovercome the effect of sublethal damage repair that occurs when twosublethal doses are separated in time.

The linear quadratic model (surviving fraction=e^(o)′u-RD2) is used tofit radiation survival data to a continuously bending curve, where D isdose and a and (3 are constants. Alpha is the linear component, ameasure of the initial slope that represents single-hit killing kineticsand dominates the radiation response at low doses. Beta is the quadraticcomponent of cell killing, that represents multiple-hit killing andcauses the curve to bend at higher doses. The alpha:beta ratio is thedose at which the linear and quadratic components of cell killing areequal. The more linear the response to killing of cells at low radiationdose, the higher is the value of alpha, and the greater is theradiosensitivity of the cells.

When the Compound of the Invention and additional anticancer treatmentmodality are administered to a subject concurrently, the term“concurrently” is not limited to the administration of a Compound of theInvention and an additional anticancer treatment modality at exactly thesame time, but rather it is meant that they are administered to asubject in a sequence and within a time interval such that they can actsynergistically to provide an increased benefit than if they wereadministered otherwise. For example, the Compounds of the Invention maybe administered at the same time or sequentially in any order atdifferent points in time as an additional anticancer treatment modality;however, if not administered at the same time, they should beadministered sufficiently close in time so as to provide the desiredtherapeutic effect, preferably in a synergistic fashion. The Compound ofthe Invention and the additional anticancer treatment modality can beadministered separately, in any appropriate form and by any suitableroute. When the Compound of the Invention and the additional anticancertreatment modality are not administered concurrently, it is understoodthat they can be administered in any order to a subject in need thereof.For example, a Compound of the Invention can be administered prior to(e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeksbefore), concomitantly with, or subsequent to (e.g., 5 minutes, 15minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks,4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) theadministration of an additional anticancer treatment modality (e.g.,radiotherapy), to a subject in need thereof. In various embodiments theCompound of the Invention and the additional anticancer treatmentmodality are administered 1 minute apart, 10 minutes apart, 30 minutesapart, less than 1 hour apart, 1 hour apart, 1 hour to 2 hours apart, 2hours to 3 hours apart, 3 hours to 4 hours apart, 4 hours to 5 hoursapart, 5 hours to 6 hours apart, 6 hours to 7 hours apart, 7 hours to 8hours apart, 8 hours to 9 hours apart, 9 hours to 10 hours apart, 10hours to 11 hours apart, 11 hours to 12 hours apart, no more than 24hours apart or no more than 48 hours apart. In one embodiment, thecomponents of the combination therapies of the invention areadministered within the same office or hospital visit. In anotherembodiment, the Compound of the Invention and the additional anticancertreatment modality are administered at 1 minute to 24 hours apart.

In one embodiment, a Compound of the Invention is administered prior orsubsequent to an additional anticancer treatment modality, preferably atleast an hour, five hours, 12 hours, a day, a week, a month, morepreferably several months (e.g., up to three months), prior orsubsequent to administration of an additional anticancer treatmentmodality.

The present invention provides methods of treating cancers comprisingthe administration of an effective amount of a Compound of the Inventionin conjunction with recognized methods of surgery, radiotherapy andchemotherapies, including, for example, chemical-based mimics ofradiotherapy whereby a synergistic enhancement of the effectiveness ofthe recognized therapy is achieved. The effectiveness of a treatment maybe measured in clinical studies or in model systems, such as a tumormodel in mice, or cell culture sensitivity assays.

The present invention provides combination therapies that result inimproved effectiveness and/or reduced toxicity. Accordingly, in oneaspect, the invention relates to the use of the Compounds of theInvention as radiosensitizers in conjunction with radiotherapy.

When the combination therapy of the invention comprises administering aCompound of the Invention are with one or more additional anticanceragents, the Compound of the Invention and the additional anticanceragents can be administered concurrently or sequentially to a subject.The agents can also be cyclically administered. Cycling therapy involvesthe administration of one or more anticancer agents for a period oftime, followed by the administration of one or more different anticanceragents for a period of time and repeating this sequentialadministration, i.e., the cycle, in order to reduce the development ofresistance to one or more of the anticancer agents of beingadministered, to avoid or reduce the side effects of one or more of theanticancer agents being administered, and/or to improve the efficacy ofthe treatment.

An additional anticancer agent may be administered over a series ofsessions; any one or a combination of the additional anticancer agentslisted below may be administered.

The present invention includes methods for treating cancer, comprisingadministering to a subject in need thereof a Compound of the Invention,and one or more additional anticancer agents or pharmaceuticallyacceptable salts thereof. The Compound of the Invention and theadditional anticancer agent(s) can act additively or synergistically.Suitable anticancer agents include, but are not limited to, gemcitabine,capecitabine, methotrexate, taxol, taxotere, mercaptopurine,thioguanine, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide,nitrosoureas, cisplatin, carboplatin, mitomycin, dacarbazine,procarbizine, etoposide, teniposide, campathecins, bleomycin,doxorubicin, idarubicin, daunorubicin, dactinomycin, plicamycin,mitoxantrone, L-asparaginase, doxorubicin, epirubicin, 5-fluorouracil(5-FU), taxanes such as docetaxel and paclitaxel, leucovorin,levamisole, irinotecan, estramustine, etoposide, nitrogen mustards,BCNU, nitrosoureas such as carmustine and lomustine, vinca alkaloidssuch as vinblastine, vincristine and vinorelbine, platinum complexessuch as cisplatin, carboplatin and oxaliplatin, imatinib mesylate,hexamethylmelamine, topotecan, tyrosine kinase inhibitors, tyrphostinsherbimycin A, genistein, erbstatin, and lavendustin A.

In one embodiment, the anti-cancer agent can be, but is not limited to,a drug listed in Table 2.

TABLE 2 Alkylating agents Nitrogen mustards: Cyclophosphamide IfosfamideTrofosfamide Chlorambucil Nitrosoureas: Carmustine (BCNU) Lomustine(CCNU) Alkylsulphonates: Busulfan Treosulfan Triazenes: DacarbazinePlatinum complexes: Cisplatin Carboplatin Oxaliplatin Plant AlkaloidsVinca alkaloids: Vincristine Vinblastine Vindesine Vinorelbine Taxoids:Paclitaxel Docetaxel DNA Topoisomerase Inhibitors Epipodophyllins:Etoposide Teniposide Topotecan 9-aminocamptothecin CamptothecinCrisnatol Mitomycins: Mitomycin C Anti-metabolites Anti-folates: DHFRinhibitors: Methotrexate Trimetrexate IMP dehydrogenase Mycophenolicacid Inhibitors: Tiazofurin Ribavirin EICAR Ribonuclotide reductaseHydroxyurea Inhibitors: Deferoxamine Pyrimidine analogs: Uracil analogs:5-Fluorouracil Floxuridine Doxifluridine Ratitrexed Cytosine analogs:Cytarabine (ara C) Cytosine arabinoside Fludarabine GemcitabineCapecitabine Purine analogs: Mercaptopurine Thioguanine DNAAntimetabolites: 3-HP 2′-deoxy-5-fluorouridine 5-HP alpha-TGDRaphidicolin glycinate ara-C 5-aza-2′-deoxycytidine beta-TGDRcyclocytidine guanazole inosine glycodialdehyde macebecin IIPyrazoloimidazole Hormonal therapies: Receptor antagonists:Anti-estrogen: Tamoxifen Raloxifene Megestrol LHRH agonists: GoserelinLeuprolide acetate Anti-androgens: Flutamide BicalutamideRetinoids/Deltoids Cis-retinoic acid Vitamin A derivative: All-transretinoic acid (ATRA-IV) Vitamin D3 analogs: EB 1089 CB 1093 KH 1060Photodynamic therapies: Vertoporfin (BPD-MA) PhthalocyaninePhotosensitizer Pc4 Demethoxy-hypocrellin A (2BA-2-DMHA) Cytokines:Interferon-α Interferon-β Interferon-γ Tumor necrosis factorAngiogenesis Angiostatin (plasminogen fragment) Inhibitors:antiangiogenic antithrombin III Angiozyme ABT-627 Bay 12-9566 BenefinBevacizumab BMS-275291 cartilage-derived inhibitor (CDI) CAI CD59complement fragment CEP-7055 Col 3 Combretastatin A-4 Endostatin(collagen XVIII fragment) Fibronectin fragment Gro-beta HalofuginoneHeparinases Heparin hexasaccharide fragment HMV833 Human chorionicgonadotropin (hCG) IM-862 Interferon alpha/beta/gamma Interferoninducible protein (IP-10) Interleukin-12 Kringle 5 (plasminogenfragment) Marimastat Metalloproteinase inhibitors (TIMPs)2-Methoxyestradiol MMI 270 (CGS 27023A) MoAb IMC-1C11 Neovastat NM-3Panzem PI-88 Placental ribonuclease inhibitor Plasminogen activatorinhibitor Platelet factor-4 (PF4) Prinomastat Prolactin 16 kD fragmentProliferin-related protein (PRP) PTK 787/ZK 222594 Retinoids SolimastatSqualamine SS 3304 SU 5416 SU6668 SU11248 Tetrahydrocortisol-STetrathiomolybdate Thalidomide Thrombospondin-1 (TSP-1) TNP-470Transforming growth factor-beta (TGF-β) Vasculostatin Vasostatin(calreticulin fragment) ZD6126 ZD 6474 farnesyl transferase inhibitors(FTI) Bisphosphonates Antimitotic agents: Allocolchicine Halichondrin BColchicine colchicine derivative dolstatin 10 Maytansine RhizoxinThiocolchicine trityl cysteine Others: Isoprenylation inhibitors:Dopaminergic 1-methyl-4-phenylpyridinium ion neurotoxins: Cell cycleinhibitors: Staurosporine Actinomycins: Actinomycin D DactinomycinBleomycins: Bleomycin A2 Bleomycin B2 Peplomycin Anthracyclines:Daunorubicin Doxorubicin (adriamycin) Idarubicin Epirubicin PirarubicinZorubicin Mitoxantrone MDR inhibitors: Verapamil Ca²⁺ATPase inhibitors:Thapsigargin

Other anti-cancer agents that may be used in the present inventioninclude, but are not limited to, acivicin; aclarubicin; acodazolehydrochloride; acronine; adozelesin; aldesleukin; altretamine;ambomycin; ametantrone acetate; aminoglutethimide; amsacrine;anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa;azotomycin; batimastat; benzodepa; bicalutamide; bisantrenehydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate;brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone;caracemide; carbetimer; carboplatin; carmustine; carubicinhydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin;cisplatin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine;dacarbazine; dactinomycin; daunorubicin hydrochloride; decitabine;dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; docetaxel;doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifenecitrate; drornostanolone propionate; duazomycin; edatrexate;eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate;epipropidine; epirubicin hydrochloride; erbulozole; esorubicinhydrochloride; estramustine; estramustine phosphate sodium; etanidazole;etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride;fazarabine; fenretinide; floxuridine; fludarabine phosphate;fluorouracil; flurocitabine; fosquidone; fostriecin sodium; gemcitabinehydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide;ilmofosine; interleukin II (including recombinant interleukin II, orrIL2), interferon alfa-2a; interferon alfa-2b; interferon alfa-n1;interferon alfa-n3; interferon beta-I a; interferon gamma-I b;iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole;leuprolide acetate; liarozole hydrochloride; lometrexol sodium;lomustine; losoxantrone hydrochloride; masoprocol; maytansine;mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate;melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium;metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin;mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride;mycophenolic acid; nocodazole; nogalamycin; ormaplatin; oxisuran;paclitaxel; pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride;semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantronehydrochloride; temoporfin; teniposide; teroxirone; testolactone;thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifenecitrate; trestolone acetate; triciribine phosphate; trimetrexate;trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracilmustard; uredepa; vapreotide; verteporfin; vinblastine sulfate;vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate;vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate;vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin;zinostatin; zorubicin hydrochloride.

Further anti-cancer drugs that can be used in the present inventioninclude, but are not limited to: 20-epi-1,25-dihydroxyvitamin D3;5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol;adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine;amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine;anagrelide; anastrozole; andrographolide; angiogenesis inhibitors;antagonist D; antagonist G; antarelix; anti-dorsalizing morphogeneticprotein-1; antiandrogen, prostatic carcinoma; antiestrogen;antineoplaston; antisense oligonucleotides; aphidicolin glycinate;apoptosis gene modulators; apoptosis regulators; apurinic acid;ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane;atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron;azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat;BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactamderivatives; beta-alethine; betaclamycin B; betulinic acid; bFGFinhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide;bistratene A; bizelesin; breflate; bropirimine; budotitane; buthioninesulfoximine; calcipotriol; calphostin C; camptothecin derivatives;canarypox IL-2; carboxamide-amino-triazole; carboxyamidotriazole; CaRestM3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinaseinhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins;chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine;clomifene analogues; clotrimazole; collismycin A; collismycin B;combretastatin A4; combretastatin analogue; conagenin; crambescidin 816;crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A;cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate;cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B;deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil;diaziquone; didemnin B; didox; diethylnorspermine;dihydro-5-azacytidine; dihydrotaxol, 9-; dioxamycin; diphenylspiromustine; docetaxel; docosanol; dolasetron; doxifluridine;droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine;edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin;epristeride; estramustine analogue; estrogen agonists; estrogenantagonists; etanidazole; etoposide phosphate; exemestane; fadrozole;fazarabine; fenretinide; filgrastim; finasteride; flavopiridol;flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;imidazoacridones; imiquimod; immunostimulant peptides; insulin-likegrowth factor-1 receptor inhibitor; interferon agonists; interferons;interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact;irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum complexes; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustardanti-cancer agent; mycaperoxide B; mycobacterial cell wall extract;myriaporone; N-acetyldinaline; N-substituted benzamides; nafarelin;nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim;nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase;nilutamide; nisamycin; nitric oxide modulators; nitroxide antioxidant;nitrullyn; O6-benzylguanine; octreotide; okicenone; oligonucleotides;onapristone; ondansetron; ondansetron; oracin; oral cytokine inducer;ormaplatin; osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxelanalogues; paclitaxel derivatives; palauamine; palmitoylrhizoxin;pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine;pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin;pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum complexes;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen binding protein; sizofiran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatinstimalamer.

It is a further aspect of the invention the Compounds of the Inventioncan be administered in conjunction with chemical agents that areunderstood to mimic the effects of radiotherapy and/or that function bydirect contact with DNA. Preferred agents for use in combination withthe Compounds of the Invention for treating cancer include, but are notlimited to cis-diamminedichloro platinum (II) (cisplatin), doxorubicin,5-fluorouracil, taxol, and topoisomerase inhibitors such as etoposide,teniposide, irinotecan and topotecan.

Additionally, the invention provides methods of treatment of cancerusing the Compounds of the Invention as an alternative to chemotherapyalone or radiotherapy alone where the chemotherapy or the radiotherapyhas proven or can prove too toxic, e.g., results in unacceptable orunbearable side effects, for the subject being treated. The subjectbeing treated can, optionally, be treated with another anticancertreatment modality such as chemotherapy, surgery, or immunotherapy,depending on which treatment is found to be acceptable or bearable.

The Compounds of the Invention can also be used in an in vitro or exvivo fashion, such as for the treatment of certain cancers, including,but not limited to leukemias and lymphomas, such treatment involvingautologous stem cell transplants. This can involve a multi-step processin which the subject's autologous hematopoietic stem cells are harvestedand purged of all cancer cells, the subject is then administered anamount of a Compound of the Invention effective to eradicate thesubject's remaining bone-marrow cell population, then the stem cellgraft is infused back into the subject. Supportive care is then providedwhile bone marrow function is restored and the subject recovers.

5.8.1.4 Other Therapeutic Agents

The present methods for treating cancer can further comprise theadministration of a Compound of the Invention and an additionaltherapeutic agent or pharmaceutically acceptable salts, solvates orhydrates thereof. In one embodiment, a composition comprising a Compoundof the Invention is administered concurrently with the administration ofone or more additional therapeutic agent(s), which may be part of thesame composition or in a different composition from that comprising theCompound of the Invention. In another embodiment, a Compound of theInvention is administered prior to or subsequent to administration ofanother therapeutic agent(s).

In the present methods for treating cancer the other therapeutic agentmay be an antiemetic agent. Suitable antiemetic agents include, but arenot limited to, metoclopromide, domperidone, prochlorperazine,promethazine, chlorpromazine, trimethobenzamide, ondansetron,granisetron, hydroxyzine, acethylleucine monoethanolamine, alizapride,azasetron, benzquinamide, bietanautine, bromopride, buclizine,clebopride, cyclizine, dimenhydrinate, diphenidol, dolasetron,meclizine, methallatal, metopimazine, nabilone, oxyperndyl, pipamazine,scopolamine, sulpiride, tetrahydrocannabinols, thiethylperazine,thioproperazine and tropisetron.

In a preferred embodiment, the anti-emetic agent is granisetron orondansetron.

In another embodiment, the other therapeutic agent may be anhematopoietic colony stimulating factor. Suitable hematopoietic colonystimulating factors include, but are not limited to, filgrastim,sargramostim, molgramostim and epoietin alfa.

In still another embodiment, the other therapeutic agent may be anopioid or non-opioid analgesic agent. Suitable opioid analgesic agentsinclude, but are not limited to, morphine, heroin, hydromorphone,hydrocodone, oxymorphone, oxycodone, metopon, apomorphine, normorphine,etorphine, buprenorphine, meperidine, lopermide, anileridine,ethoheptazine, piminidine, betaprodine, diphenoxylate, fentanil,sufentanil, alfentanil, remifentanil, levorphanol, dextromethorphan,phenazocine, pentazocine, cyclazocine, methadone, isomethadone andpropoxyphene. Suitable non-opioid analgesic agents include, but are notlimited to, aspirin, celecoxib, rofecoxib, diclofinac, diflusinal,etodolac, fenoprofen, flurbiprofen, ibuprofen, ketoprofen, indomethacin,ketorolac, meclofenamate, mefanamic acid, nabumetone, naproxen,piroxicam and sulindac.

In still another embodiment, the other therapeutic agent may be ananxiolytic agent. Suitable anxiolytic agents include, but are notlimited to, buspirone, and benzodiazepines such as diazepam, lorazepam,oxazapam, chlorazepate, clonazepam, chlordiazepoxide and alprazolam.

5.8.2 Treatment of Neurological Diseases

Neurological diseases can be treated or prevented by administration ofamounts of the Compounds of the invention that are effective to treatthe neurological disease or by administration of a pharmaceuticalcomposition comprising amounts of the Compounds of the invention thatare effective to treat the neurological disease. In one embodiment, theneurological diseases that can be treated or prevented by administeringa Compound of the Invention include, but are not limited to,Huntington's disease, lupus, schizophrenia, multiple sclerosis, musculardystrophy, drug-induced movement disorders, Creutzfeldt-Jakob disease,amyotrophic lateral sclerosis, Pick's disease, Alzheimer's disease, Lewybody dementia, cortico basal degeneration, dystonia, myoclonus,Tourette's Syndrome, tremor, chorea, restless leg syndrome, Parkinson'sdisease, and Parkinsonian Syndromes, such as progressive supranuclearpalsy, multiple system atrophy, Wilson's disease and mult-infarct state.In a preferred embodiment, the neurological disease treated isHuntingon's disease, lupus, or schizophrenia.

The present invention is not to be limited in scope by the specificembodiments disclosed in the examples which are intended asillustrations of a few aspects of the invention and any embodiments thatare functionally equivalent are within the scope of this invention.Indeed, various modifications of the invention in addition to thoseshown and described herein will become apparent to those skilled in theart and are intended to fall within the scope of the appended claims.

A number of references have been cited throughout this specification,the entire disclosures of each of which have been incorporated herein byreference in their entireties for all purposes.

6. EXAMPLES 6.1 Example 1 Preparation of4-[3-(4-Dimethylamino-benzyl)-ureido]-N-hydroxy-butyramide (2)

4-[3-(4-Dimethylamino-benzyl)-ureido]-butyric acid benzyl ester

To a vigorous stirred suspension of 4-amino-butyric acid benzyl estertoluene-4-sulfonic acid (0330 g, 2.00 mmol) and triphosgene (0.200 g,0.667 mmol) in dichloromethane (40 mL) at −78° C. under nitrogenatmosphere, was added triethylamine (1.0 mL, 7.188 mmol, in 10 mLdichloromethane) dropwise via an additional funnel over a period of 2hours. The cooling bath was removed and the resulting reaction wasallowed to stir at room temperature for 1 hour, after which time4-dimethylaminobenzylamine dihydrochloride (0.446 g, 2.00 mmol) wasadded to the reaction mixture, followed by triethylamine (0.67 mL, 4.8mmol). The resulting reaction was allowed to stir for about 18 h, thenthe reaction mixture was diluted with brine (20 mL), transferred to aseparatory funnel and the aqueous layer was extracted withdichloromethane. The organic layer was dried over sodium sulfate andconcentrated in vacuo to provide a crude residue which was purified byflash column chromatography (silica gel 60, CH₂Cl₂/EtOAc=4:1 to 2:1) toafford 4-[3-(4-Dimethylamino-benzyl)-ureido]-butyric acid benzyl ester(0.686 g, 93% yield). ¹H NMR (300 MHz, CDCl₃) δ (ppm) 7.31 (m, 5H), 7.09(d, J=9.0 Hz, 2H), 6.63 (d, J=9.0 Hz, 2H), 5.20 (t, J=5.4 Hz, 1H), 5.09(t, J=5.4 Hz, 1H), 5.05 (s, 2H), 4.14 (d, J=5.7 Hz, 2H), 3.09 (dt,J=6.6, 6.3 Hz, 2H), 2.86 (s, 6H), 2.32 (t, J=7.5 Hz, 2H), 1.73 (m, 2H).¹³C NMR (75 MHz, CDCl₃) δ (ppm) 173.3, 158.5, 149.8, 135.8, 128.5,128.15, 128.09, 127.0, 112.6, 66.2, 43.9, 40.7, 39.5, 31.5, 25.4.

N-Benzyloxy-4-[3-(4-dimethylamino-benzyl)-ureido]-butyramide

To a solution of 4-[3-(4-dimethylamino-benzyl)-ureido]-butyric acidbenzyl ester (626 mg, 1.696 mmol) in methanol (20 mL) was added 10%palladium on carbon (40 mg), and the resulting reaction was allowed tostir under a hydrogen atmosphere for 18 hr, after which time thereaction mixture was filtered and concentrated in vacuo to provide acrude residue.

To a suspension of the crude residue in dichloromethane (40 mL) wasadded EDCI (650 mg, 3.391 mmol) at 0° C., followed by the benzyloxyaminehydrochloride (541 mg, 3.389 mmol) and triethylamine (0.475 mL, 3.414mmol). The cooling bath was removed and the resulting reaction wasallowed to stir at room temperature for 18 h. The reaction mixturetransferred to a separatory funnel, washed with brine (20 mL), and theaqueous layer was back extracted with dichloromethane. The combinedorganic extracts were dried over sodium sulfate and concentrated invacuo to provide a crude residue which was purified using flash columnchromatography (silica gel 60, CH₂Cl₂/MeOH=50:1 to 20:1) to provideN-Benzyloxy-4-[3-(4-dimethylamino-benzyl)-ureido]-butyramide (330 mg,50% yield). ¹H NMR (300 MHz, CDCl₃) δ (ppm) 9.98 (br s, 1H), 7.35 (m,5H), 7.11 (d, J=8.7 Hz, 2H), 6.65 (d, J=8.7 Hz, 2H), 5.05 (br s, 2H),4.86 (s, 2H), 4.16 (d, J=5.4 Hz, 2H), 3.09 (dt, J=6.6, 6.0 Hz, 2H), 2.89(s, 6H), 2.02 (t, J=6.6 Hz, 2H), 1.68 (m, 2H). ¹³C NMR (75 MHz, CDCl₃) δ(ppm) 171.1, 159.0, 150.0, 129.1, 128.6, 128.5, 126.9, 112.8, 78.0,44.1, 40.7, 38.9, 30.3, 26.8.

4-[3-(4-Dimethylamino-benzyl)-ureido]-N-hydroxy-butyramide (2)

To a solution of theN-benzyloxy-4-[3-(4-dimethylamino-benzyl)-ureido]-butyramide (287 mg,0.747 mmol) in methanol (15 mL) was added 10% palladium on carbon (30mg) and the resulting reaction was allowed to stir under a hydrogenatmosphere for 18 hr, after which time the reaction mixture was filteredand concentrated in vacuo to provide compound 2, which was used withoutfurther purification. (203 mg, 92% yield). ¹H NMR (300 MHz, DMSO-d₆) δ(ppm) 10.38 (s, 1H), 8.69 (s, 1H), 7.06 (d, J=9.0 Hz, 2H), 6.67 (d,J=9.0 Hz, 2H), 6.10 (t, J=5.4 Hz, 1H), 5.87 (t, J=5.4 Hz, 1H), 4.06 (d,J=5.7 Hz, 2H), 2.97 (dt, J=6.6, 6.3 Hz, 2H), 2.85 (s, 6H), 1.94 (t,J=7.2 Hz, 2H), 1.57 (m, 2H). ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm) 168.8,157.9, 149.4, 128.2, 127.9, 112.3, 42.4, 40.2, 38.8, 29.8, 26.3.

6.2 Example 2 Preparation of5-[3-(4-dimethylamino-benzyl)-ureido]-pentanoic acid hydroxyamide (3)

Compound 3 was prepared using the methodology described for thepreparation of compound 2, by substituting 4-amino-butyric acid benzylester toluene-4-sulfonic acid with 5-amino-pentanoic acid benzyl estertoluene-4-sulfonic acid. ¹H NMR (300 MHz, DMSO-d₆) δ (ppm) 10.35 (br s,1H), 8.68 (br s, 1H), 7.06 (d, J=8.7 Hz, 2H), 6.67 (d, J=8.7 Hz, 2H),6.07 (t, J=5.4 Hz, 1H), 5.84 (t, J=5.4 Hz, 1H), 4.05 (d, J=6.0 Hz, 2H),2.97 (dt, J=6.6, 6.3 Hz, 2H), 2.84 (s, 6H), 1.94 (t, J=7.2 Hz, 2H), 1.47(m, 2H), 1.32 (m, 2H). ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm) 168.7, 157.7,149.2, 128.0, 127.7, 112.1, 42.2, 40.0, 38.6, 31.7, 29.4, 22.3.

6.3 Example 3 Preparation of6-[3-(4-dimethylamino-benzyl)-ureido]-hexanoic acid hydroxyamide (4)

Compound 4 was prepared using the methodology described for thepreparation of compound 2, by substituting 4-amino-butyric acid benzylester toluene-4-sulfonic acid with 6-amino-hexanoic acid benzyl estertoluene-4-sulfonic acid. ¹H NMR (300 MHz, DMSO-d₆) δ (ppm) 10.34 (br, s,1H), 8.67 (br s, 1H), 7.06 (d, J=8.7 Hz, 2H), 6.67 (d, J=8.7 Hz, 2H),6.05 (t, J=5.7 Hz, 1H), 5.80 (t, J=5.7 Hz, 1H), 4.05 (d, J=6.3 Hz, 2H),2.97 (dt, J=6.6, 6.3 Hz, 2H), 2.85 (s, 6H), 1.93 (t, J=7.5 Hz, 2H), 1.48(m, 2H), 1.34 (m, 2H), 1.22 (m, 2H). ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm)168.7, 157.7, 149.2, 128.0, 127.7, 112.1, 42.2, 40.0, 38.9, 31.9, 29.5,25.7, 24.6.

6.4 Example 4 Preparation of7-[3-(4-dimethylamino-benzyl)-ureido]-heptanoic acid hydroyxamide (5)

Compound 5 was prepared using the methodology described for thepreparation of compound 2, by substituting 4-amino-butyric acid benzylester toluene-4-sulfonic acid with 7-amino-heptanoic acid benzyl estertoluene-4-sulfonic acid. ¹H NMR (300 MHz, DMSO-d₆) δ ppm) 10.33 (br, s,1H), 8.66 (br s, 1H), 7.05 (d, J=8.7 Hz, 2H), 6.67 (d, J=8.7 Hz, 2H),6.04 (t, J=5.7 Hz, 1H), 5.79 (t, J=5.7 Hz, 1H), 4.05 (d, J=6.0 Hz, 2H),2.97 (dt, J=6.6, 6.6 Hz, 2H), 2.84 (s, 6H), 1.93 (t, J=7.5 Hz, 2H), 1.47(m, 2H), 1.34 (m, 2H), 1.22 (m, 4H)

6.5 Example 5 Preparation of8-[3-(4-dimethylamino-benzyl)-ureido]-octanoic acid hydroxyamide (6)

Compound 6 was prepared using the methodology described for thepreparation of compound 2, by substituting 4-amino-butyric acid benzylester toluene-4-sulfonic acid with 8-amino-octanoic acid benzyl estertoluene-4-sulfonic acid. ¹H NMR (300 MHz, DMSO-d₆) δ (ppm) 10.33 (br, s,1H), 8.66 (br s, 1H), 7.05 (d, J=8.4 Hz, 2H), 6.67 (d, J=8.4 Hz, 2H),6.05 (t, J=5.7 Hz, 1H), 5.81 (t, J=5.7 Hz, 1H), 4.05 (d, J=6.0 Hz, 2H),2.97 (dt, J=6.6, 6.0 Hz, 2H), 2.85 (s, 6H), 1.93 (t, J=7.5 Hz, 2H), 1.47(m, 2H), 1.34 (m, 2H), 1.24 (m, 6H).

6.6 Example 6 Preparation of7-[3-(4-dimethylamino-phenyl)-ureido]-heptanoic acid hydroxyamide (7)

Compound 7 was prepared using the methodology described for thepreparation of compound 2, by substituting 4-amino-butyric acid benzylester toluene-4-sulfonic acid with 7-amino-heptanoic acid benzyl estertoluene-4-sulfonic acid and 4-dimethylaminobenzylamine dihydrochloridewith N,N-dimethyl-benzene-1,4-diamine dihydrochloride. ¹H NMR (300 MHz,DMSO-d₆) δ (ppm) 10.34 (br, s, 1H), 8.64 (br s, 1H), 7.98 (br s, 1H),7.17 (d, J=9.0 Hz, 2H), 6.65 (d, J=9.0 Hz, 2H), 5.91 (t, J=5.7 Hz, 1H),3.03 (dt, J=6.6, 6.0 Hz, 2H), 2.79 (s, 6H), 1.94 (t, J=7.5 Hz, 2H), 1.48(m, 2H), 1.39 (m, 2H), 1.25 (m, 4H).

6.7 Example 7 Preparation of 6-(3-adamantan-1-yl-ureido)-hexanoic acidhydroxyamide (8) 6-(3-Adamantan-1-yl-ureido)-hexanoic acid benzyl ester

To a vigorously stirred suspension of 6-amino-hexanoic acid benzyl estertoluene-4-sulfonic acid (0.786 g, 2.00 mmol) and triphosgene (0.200 g,0.667 mmol) in dichloromethane (30 mL) at −78° C. under inertatmosphere, was added triethylamine (1.0 mL, 7.188 mmol, in 10 mLdichloromethane) dropwise via additional funnel over a period of 2hours. The cooling bath was removed and the resulting reaction wasallowed to stir at room temperature for 1 hour, after which time,4-Dimethylaminobenzylamine dihydrochloride (0.303 g, 2.00 mmol) wasadded to the reaction mixture. After stirring overnight, the mixture waswashed with brine (20 mL) and the aqueous layer was extracted withdichloromethane. The combined organic layers were dried over sodiumsulfate and the solvent was evaporated. The residue was purified usingflash column chromatography (silica gel 60, CH₂Cl₂/EtOAc=6:1 to 4:1) toafford 6-(3-Adamantan-1-yl-ureido)-hexanoic acid benzyl ester 0.481 g(61% yield). ¹H NMR (300 MHz, CDCl₃) δ (ppm) 7.34 (m, 5H), 5.11 (s, 2H),4.61 (t, J=5.7 Hz, 1H), 4.43 (s, 1H), 3.08 (dt, J=6.9, 5.7 Hz, 2H), 2.35(t, J=7.5 Hz, 2H), 2.04 (m, 3H), 1.94 (m, 6H), 1.65 (m, 8H) 1.46 (m, 2H)1.31 (m, 2H). ¹³C NMR (75 MHz, CDCl₃) δ (ppm) 173.5, 157.4, 135.9,128.5, 128.2, 128.1, 66.1, 50.6, 42.5, 39.8, 36.4, 34.1, 29.9, 29.5,26.4, 24.5.

6-(3-Adamantan-1-yl-ureido)-hexanoic acid hydroxyamide (8)

To a solution of 6-(3-adamantan-1-yl-ureido)-hexanoic acid benzyl ester(440 mg, 1.696 mmol) in methanol (20 mL) was added 10% palladium oncarbon (40 mg) and the resulting reaction was allowed to stir under ahydrogen atmosphere for 18 hr, after which time the reaction mixture wasfiltered and concentrated in vacuo to provide a crude residue (343 mg).

To a suspension of the crude residue in dichloromethane (40 mL) wasadded EDCI (427 mg, 2.23 mmol) at 0° C., followed by the addition ofbenzyloxyamine hydrochloride (267 mg, 1.67 mmol) and triethylamine (0.23mL, 1.67 mmol). The cooling bath was removed and the mixture was allowedto stir at room temperature overnight. The resulting reaction wasallowed to stir for about 18 h, then the reaction mixture was dilutedwith brine (20 mL), transferred to a separatory funnel and the aqueouslayer was extracted with dichloromethane. The organic layer was driedover sodium sulfate and concentrated in vacuo to provide a crude residuewhich was purified by flash column chromatography to provide amide (262mg).

To a solution of the amide in methanol (15 mL) was added 10% palladiumon carbon (30 mg). After it was treated with hydrogen under atmospherepressure overnight, the reaction mixture was filtered and concentratedto provide compound 8 (184 mg, 51% yield from6-(3-adamantan-1-yl-ureido)-hexanoic acid benzyl ester). ¹H NMR (300MHz, CD₃OD) δ (ppm) 3.04 (t, J=7.2 Hz, 2H), 2.09 (t, J=7.5 Hz, 2H), 2.03(m, 3H), 1.96 (m, 6H), 1.70 (m, 6H), 1.62 (m, 2H), 1.45 (m, 2H), 1.34(m, 2H). ¹³C NMR (75 MHz, CD₃OD) δ (ppm) 173.0, 160.5, 51.5, 43.6, 40.5,37.7, 33.8, 31.19, 31.14, 27.5, 26.6.

6.8 Example 8 Preparation of2-mercapto-N-[5-(3-phenyl-ureido)-pentyl]-acetamide (9)

Tritylsulfanyl-acetic acid methyl ester

To a mixture of methyl mercaptoacetate (5.30 g, 50 mmol) andtriphenylmethanol (13.0 g, 50 mmol) in chloroform (20 mL) was addedtrifloroacetic acid (5 mL) in 5 min. After stirring at room temperaturefor 1 h, the volatiles were removed in vacuo. The crude product waspurified by recrystallization (dichloromethane/Hexane=1/2) to provideTritylsulfanyl-acetic acid methyl ester (15.9 g, 91%). ¹H NMR (300 MHz,CDCl₃) δ 7.44-7.38 (m, 6H), 7.34-7.18 (m, 9H), 3.58 (s, 3H), 2.98 (s,2H).

7-Amino-1-tritylsulfanyl-heptan-2-one

1,5-Diaminopentane (0.75 g, 7.21 mmol) was stirred whiletritylsulfanyl-acetic acid methyl ester (2.53 g, 7.27 mmol) was addedslowly. The mixture was heated at 100° C. for 2 h while methyl alcoholescaped. The product was isolated by column chromatography(dichloromethane/MeOH/Et₃N=10/1/0.1) to provide7-Amino-1-tritylsulfanyl-heptan-2-one (1.63 g, 54%). ¹H NMR (300 MHz,CD₃OD) δ 7.42-7.36 (m, 6H), 7.31-7.18 (m, 9H), 3.00 (t, J=6.9 Hz, 2H),2.60 (t, J=6.9 Hz, 2H), 1.42 (m, 4H), 1.27 (m, 2H). ¹³C NMR (75 MHz,CD₃OD) δ 171.1, 145.8, 130.9, 129.2, 128.2, 68.4, 42.5, 40.8, 37.4,33.4, 30.1, 25.3.

2-Mercapto-N-[5-(3-phenyl-ureido)-pentyl]-acetamide (9)

To a solution of 7-amino-1-tritylsulfanyl-heptan-2-one (0.36 g, 0.86mmol) in dichloromethane (10 mL) was added phenylisocyanate (0.10 ml,0.92 mmol). The mixture was stirred at room temperature for 2 h. Solventwas then removed in vacuo and the crude product was purified by columnchromatography (dichloromethane/MeOH=100/1 to 40/1) to provide1-(6-Oxo-7-tritylsulfanyl-heptyl)-3-phenyl-urea (0.43 g, 93%).

To a solution of 1-(6-oxo-7-tritylsulfanyl-heptyl)-3-phenyl-urea (0.420g, 0.78 mmol) in dichloromethane was added trifluoroacetic acid (0.50mL, 6.49 mmol), followed by the addition of triethylsilane (0.18 mL,1.13 mmol). After stirring at room temperature for 2 h, the volatileswere removed in vacuo and the crude product was purified by columnchromatography (dichloromethane/MeOH=60/1 to 20/1) to provide compound 9(0.216 g, 94%). ¹H NMR (300 MHz, DMSO-d₆) δ (ppm) 8.39 (br, s, 1H), 7.99(br t, J=5.1 Hz, 1H), 7.38 (d, J=7.5 Hz, 2H), 7.21 (d, J=7.5 Hz, 2H),6.87 (t, J=7.5 Hz, 1H), 6.10 (br s, 1H), 3.08 (d, J=7.8 Hz, 2H), 3.05(t, J=6.9 Hz, 2H), 2.71 (t, J=7.8 Hz, 1H), 1.43 (m, 4H), 1.28 (m, 2H).¹³C NMR (75 MHz, DMSO-d₆) δ (ppm) 169.4, 155.2, 140.6, 128.6, 120.9,117.6, 38.9, 38.8, 29.5, 28.7, 27.2, 23.8.

6.9 Example 9 Preparation of2-Mercapto-N-[6-(3-phenyl-ureido)-hexyl]-acetamide (10)

Compound 10 was prepared using the methodology described for thepreparation of compound 9, by substituting 1,5-diaminopentane with1,6-diaminohexane. ¹H NMR (300 MHz, DMSO-d₆) δ (ppm) 8.37 (br s, 1H),7.97 (br t, J=5.1 Hz, 1H), 7.37 (d, J=7.5 Hz, 2H), 7.20 (t, J=7.5 Hz,2H), 6.87 (t, J=7.5 Hz, 1H), 6.10 (br t, J=5.4 Hz, 1H), 3.07 (d, J=7.8Hz, 2H), 3.04 (t, J=6.9 Hz, 2H), 2.71 (t, J=7.8 Hz, 1H), 1.41 (m, 4H),1.28 (m, 4H). ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm) 169.4, 155.2, 140.6,128.6, 120.9, 117.5, 38.96, 38.79, 29.7, 29.0, 27.1, 26.1 (2C).

6.10 Example 10 Preparation ofN-[5-(3-benzyl-ureido)-pentyl]-2-mercapto-acetamide (11)

Compound 11 was prepared using the methodology described for thepreparation of Compound 9, by substituting phenylisocyanate withbenzylisocyanate. ¹H NMR (300 MHz, DMSO-d₆) δ (ppm) 7.98 (br s, 1H),7.35-7.18 (m, 5H), 6.27 (br s, 1H), 5.91 (br s, 1H), 4.19 (s, 2H), 3.07(d, J=8.1 Hz, 2H), 3.02 (m, 4H), 2.71 (t, J=8.1 Hz, 1H), 1.37 (m, 4H),1.26 (m, 2H). ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm) 169.4, 158.1, 141.0,128.2, 127.0, 126.5, 42.9, 39.2, 38.8, 29.7, 28.7, 27.1, 23.7.

6.11 Example 11 Preparation ofN-{6-[3-(4-dimethylamino-benzyl)-ureido]-hexyl}-2-mercapto-acetamide(12)

Compound 12 was prepared using the methodology described for thepreparation of compound 9, by substituting 1,5-diaminopentane with1,6-diaminohexane and phenylisocyanate with(4-isocyanato-phenyl)-dimethyl-amine. ¹H NMR (300 MHz, DMSO-d₆) δ (ppm)7.98 (br s, 1H), 7.05 (d, J=8.7 Hz, 2H), 6.60 (d, J=8.7 Hz, 2H), 6.04(br t, J=5.7 Hz, 1H), 5.80 (br t, J=5.7 Hz, 1H), 4.05 (d, J=5.7 Hz, 2H),3.06 (d, J=8.1 Hz, 2H), 3.04 (dt, J=6.9, 6.0 Hz, 2H), 2.98 (dt, J=6.6,6.0 Hz, 2H), 2.84 (s, 6H), 2.71 (t, J=8.1 Hz, 1H), 1.36 (m, 4H), 1.24(m, 4H). ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm) 169.2, 157.9, 149.3, 127.9,126.9, 112.3, 42.4, 40.3, 39.1, 38.7, 30.0, 29.0, 27.1, 26.12, 26.08.

6.12 Example 12 Preparation of2-mercapto-N-[6-(3-phenyl-ureido)-hexyl]-acetamide (13)

Compound 13 was prepared using the methodology described for thepreparation of compound 9, by substituting 1,5-diaminopentane with1,6-diaminohexane. ¹H NMR (300 MHz, DMSO-d₆) δ (ppm) 8.37 (br s, 1H),7.97 (br t, J=5.1 Hz, 1H), 7.37 (d, J=7.5 Hz, 2H), 7.20 (t, J=7.5 Hz,2H), 6.87 (t, J=7.5 Hz, 1H), 6.10 (br t, J=5.4 Hz, 1H), 3.07 (d, J=7.8Hz, 2H), 3.04 (t, J=6.9 Hz, 2H), 2.71 (t, J=7.8 Hz, 1H), 1.41 (m, 4H),1.28 (m, 4H). ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm) 169.4, 155.2, 140.6,128.6, 120.9, 117.5, 38.96, 38.79, 29.7, 29.0, 27.1, 26.1 (2C).

6.13 Example 13 Preparation ofN-hydroxy-3-[3-(2-hydroxy-2-phenyl-ethyl)-ureido]-propionamide (14)

Compound 14 was prepared using the methodology described for thepreparation of Compound 2, by substituting 4-amino-butyric acid benzylester toluene-4-sulfonic acid with 3-amino-propionic acid benzyl estertoluene-4-sulfonic acid and 4-dimethylaminobenzylamine dihydrochloridewith 2-amino-1-phenyl-ethanol. ¹H NMR (CD₃OD) δ 7.40-7.22 (m, 5H), 4.69(dd, J=7.8, 4.2 Hz, 1H), 3.41-3.34 (m, 3H), 3.22 (dd, J=13.8, 7.8 Hz,1H), 2.25 (t, J=6.9 Hz, 2H). ¹³C NMR (CD₃OD) δ 171.1, 161.3, 144.2,129.5, 128.7, 127.3, 74.6, 48.9, 37.5, 34.7.

6.14 Example 14 Preparation ofN-hydroxy-4-[3-(2-hydroxy-2-phenyl-ethyl)-ureido]-butyramide (15)

Compound 15 was prepared using the methodology described for thepreparation of compound 2, by substituting 4-dimethylaminobenzylaminedihydrochloride with 2-amino-1-phenyl-ethanol. ¹H NMR (CD₃OD) δ7.42-7.24 (m, 5H), 4.73 (dd, J=7.8, 4.2 Hz, 1H), 3.43 (dd, J=13.8, 4.2Hz, 1H), 3.26 (dd, J=13.8, 7.8 Hz, 1H), 3.14 (t, J=6.9 Hz, 2H), 2.12 (t,J=7.2 Hz, 2H), 1.76 (m, 2H). ¹³C NMR (CD₃OD) δ 172.7, 161.5, 144.2,129.5, 128.7, 127.3, 74.6, 48.1, 39.4, 30.6, 27.0.

6.15 Example 15 Preparation ofN-Hydroxy-4-(3-phenethyl-ureido)-butyramide (1)

Compound 1 was prepared using the methodology described for thepreparation of compound 2, by substituting 4-dimethylaminobenzylaminedihydrochloride with phenethylamine. ¹H NMR (CD₃OD) δ 7.30-7.15 (m, 5H),3.34 (t, J=6.9 Hz, 2H), 3.11 (t, J=7.2 Hz, 2H), 2.76 (t, J=7.2 Hz, 2H),2.09 (t, J=7.5 Hz, 2H), 1.73 (m, 2H). ¹³C NMR (CD₃OD) δ 172.7, 161.3,140.9, 130.0, 129.6, 127.4, 42.8, 40.4, 37.7, 31.3, 27.8.

6.16 Example 16 Preparation of octanedioic acid adamantan-1-ylamidehydroxyamide (17)

7-(Adamantan-1-yl-carbamoyl)-heptanoic acid methyl ester

To a solution of oxonane-2,9-dione (323 mg, 2.071 mmol) in THF (25 mL)was added 1-adamantanamine (312 mg, 2.066 mmol). The resulting reactionwas allowed to stir at room temperature for 16 hours, then concentratedin vacuo to provide a crude residue which was diluted with methanol (20mL) and treated with AG 50 W X-2 acid resin (60 mg). The resultingreaction was heated at reflux with stirring for 5 hours, then thereaction mixture was cooled to room temperature, filtered andconcentrated in vacuo. The resulting residue was purified using flashcolumn chromatography (silica gel 60, CH₂Cl₂/EtOAc=5:1) to provide7-(adamantan-1-yl-carbamoyl)-heptanoic acid methyl ester as a whitesolid (516 mg, 78% yield from 1-adamantanamine 5). ¹H NMR (300 MHz,CDCl₃) δ 5.16 (br s, 1H), 3.66 (s, 3H), 2.30 (t, J=7.5 Hz, 2H), 2.07 (m,5H), 1.99 (m, 6H), 1.67 (m, 6H), 1.67 (m, 6H), 1.61 (m, 4H), 1.32 (m,4H). ¹³C NMR (75 MHz, CDCl₃) δ 174.2, 172.1, 51.7, 51.4, 41.6, 37.6,36.3, 34.0, 29.4, 28.8, 28.7, 25.5, 24.7.

Octanedioic acid adamantan-1-ylamide hydroxyamide (17)

To a first solution of hydroxylamine hydrochloride (66 mg, 0.950 mmol)and phenolphthalein (0.5 mg) in methanol (3 mL), was added dropwise asecond solution of sodium metal (33 mg, 1.435 mmol) in methanol (3 mL)via additional funnel until a pink endpoint was reached and precipitateappeared. To the reaction mixture was added a solution of7-(adamantan-1-yl-carbamoyl)-heptanoic acid methyl ester (152 mg, 0.474mmol) in methanol (4 mL) was added, followed by the remainder of thesecond solution of sodium metal in methanol. The resulting reaction wasallowed to stir for 24 hours, then the reaction mixture was diluted withwater (15 mL), followed by glacial acetic acid (0.2 mL) with stirring.The resulting precipitate was suction filtered and rinsed using water,then dried at room temperature under vacuum to provide a crude residuewhich was purified by recrystallization from dichloromethane/hexane toprovide Compound 17 (104 mg, 68% yield). ¹H NMR (300 MHz, CD₃OD) δ 7.31(br s, 1H), 2.09 (t, J=7.5 Hz, 2H), 2.08-1.98 (m, 11H), 1.71 (m, 6H),1.59 (m, 4H), 1.33 (m, 4H). ¹³C NMR (75 MHz, CD₃OD) δ 175.7, 173.1,52.8, 42.5, 38.0, 37.7, 33.9, 31.1, 30.00, 29.96, 27.2, 26.8.

6.17 Example 17 Preparation of octanedioic acid hydroxyamide[2-(7-hydroxycarbamoyl-heptanoylamino)-phenyl]-amide (18)

Compound 18 was prepared using the methodology described for thepreparation of compound 17, by substituting 1-adamantanamine withbenzene-1,2-diamine. ¹H NMR (300 MHz, CD₃OD) δ 7.46 (m, 2H), 7.22 (m,2H), 2.41 (t, J=7.5, 2H), 2.10 (t, J=7.5 Hz, 2H), 1.71 (m, 4H), 1.64 (m,4H), 1.40 (m, 8H). ¹³C NMR (75 MHz, CD₃OD) δ 175.0, 173.0, 132.2, 127.2,126.7, 37.6, 33.7, 29.98, 29.89, 26.77, 26.64.

6.18 Example 18 Preparation ofN-[6-(3-phenyl-ureido)-hexyl]-2-{[6-(3-phenyl-ureido)-hexylcarbamoyl]-methyldisulfanyl}-acetamide(19)

To a solution of 2-mercapto-N-[6-(3-phenyl-ureido)-hexyl]-acetamide (130mg) in CH₂Cl₂ (10 mL) and MeOH (2 mL) was added Et₃N (0.1 mL). Oxygenwas then bubbled through the resulting solution for 3 h with vigorousstirring. The reaction mixture was concentrated in vacuo and theresulting crude residue was purified using flash column chromatography(CH₂Cl₂/MeOH=60/2-60/4) to provide Compound 19 (122 mg, 94%). ¹H NMR(300 MHz, DMSO-d₆) δ (ppm) 8.36 (br s, 2H), 8.07 (br t, J=5.4 Hz, 2H),7.37 (d, J=7.5 Hz, 4H), 7.20 (t, J=7.5 Hz, 4H), 6.87 (t, J=7.5 Hz, 2H),6.10 (br t, J=5.4 Hz, 2H), 3.46 (s, 4H), 3.07 (m, 8H), 1.41 (m, 8H),1.28 (m, 8H). ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm) 167.6, 155.2, 140.6,128.6, 120.9, 117.5, 42.0, 38.97, 38.89, 29.7, 29.0, 26.13, 26.10.

6.19 Example 19 Preparation ofN-{6-[3-(4-dimethylamino-benzyl)-ureido]-hexyl}-2-({6-[3-(4-dimethylamino-benzyl)-ureido]-hexylcarbamoyl}-methyldisulfanyl)-acetamide(20)

Compound 20 was prepared using the methodology described for thepreparation of compound 19, by substituting2-mercapto-N-[6-(3-phenyl-ureido)-hexyl]-acetamide withN-{6-[3-(4-dimethylamino-benzyl)-ureido]-hexyl}-2-mercapto-acetamide. ¹HNMR (300 MHz, DMSO-d₆) δ (ppm) 8.07 (br t, J=5.4 Hz, 2H), 7.05 (d, J=8.4Hz, 4H), 6.60 (d, J=8.4 Hz, 4H), 6.05 (br t, J=6.0 Hz, 2H), 5.81 (br t,J=5.7 Hz, 2H), 4.05 (d, J=5.7 Hz, 4H), 3.45 (s, 4H), 3.06 (dt, J=6.6,6.0 Hz, 4H), 2.97 (dt, J=6.3, 6.0 Hz, 4H), 2.84 (s, 12H), 1.45-1.18 (m,16H). ¹³C NMR (75 MHz, DMSO-d₆) δ (ppm) 167.4, 157.9, 149.3, 127.9,126.9, 112.3, 42.4, 41.8, 40.3, 39.1, 38.7, 29.9, 28.9, 26.01, 25.95.

6.20 Example 20 Inhibition of HDAC Activity Assay

The HDAC activity inhibition assay was performed as follows, with datafor selected compounds being listed in Table 3: Nuclear extracts fromHeLa cells were prepared in 0.1 M KCl, 20 mM HEPES/NaOH at pH 7.9, 20%glycerol, 0.2 mM DTA, 0.5 mM DTT, and 0.5 mM PMSF (J. D. Dignam et al.Nuc. Acids Res 11:1475, 1983). Nuclear extract were mixed with Fluor deLys substrate and the indicated concentrations of the Compounds of theInvention at 37° C. in HDAC assay buffer containing 25 mM Tris/Cl, pH8.0, 137 mM NaCl, 2.7 mM KCl, and 1 mM MgCl₂. The resulting reactionswere quenched after 15 min via the addition of Fluor de Lys Developerand fluorescence was measured at an excitation wavelength of 360 nm anda detection of emitted light of 460 nm (TECAN ULTRA 384). For each testsample the corresponding assay reaction was performed in triplicate.Test samples include a Blank sample (no enzyme), a Control sample (noinhibitor), a negative control (MD83A), positive controls (TSA andSAHA), and selected Compounds of the Invention. For the selectedCompounds of the Invention, samples at the following concentrations wereprepared and tested: 1 μM, 5 μM, 10 μM, 100 μM and 1 mM. TSA and SAHAwere used at concentrations of 0.5-5 μM. M83A was used at its IC₅₀concentration (3 μM).

Results for selected compounds of the invention in the HDAC activityinhibition assay are presented in Table 3 (for 50% inhibition of HDACactivity) and in FIG. 1.

TABLE 3 HDAC Activity Inhibition Assay Data For Selected CompoundsCytotoxicity in Compound 50% HDAC SQ-20B cells Compound No. activityinhibition (IC₅₀ )

TSA 300 nM 200 nM

SAHA 700 nM 3 μM

 2 800 nM 50 μM

 3 800 nM 25 μM

 5 1 μM 50 μM

 6 700 nM 50 μM

 7 700 nM 10 μM

 8 800 nM 50 μM

 9 900 nM NA

10 900 nM 50 μM

11 1.6 μM NA

21 1.6 μM NA

22 800 nM NA

23 750 nM NA

24 800 nM NA

25 1 μM NA

6.21 Example 21 Determination of Cytotoxicity in SQ-20B Cells

To determine cytotoxicities of the Compounds of the Invention, humansquamous carcinoma cells (SQ-20B), which exhibit a radiation resistantphenotype, were treated with the Compounds of the Invention at 0, 10 μM,50 μM, 100 μM, 300 μM, 500 μM and 1 mM. For each concentrationindicated, individual T25 flasks were seeded with the following numberof SQ-20B cells: for no drug, 10 μM and 50 μM drug concentrations,separate T25 flasks were each seeded with 100 cells; for 100 μM and 300μM drug concentrations, separate T25 flasks were each seeded with 200cells each; and for 500 μM and 1 mM drug concentrations, separate T25flasks were each seeded with 300 cells each. A Levy Hemacytometer(Hausser Scientific) was used to count the cells in stock suspension.Serial dilutions of stock suspension were performed to obtain the properconcentration for cell seeding. To conduct the cytotoxicity study theSQ20B cells were first seeded under the appropriate treatmentspecifications and allowed to settle for 24 hr in a tissue cultureincubator set at 37° C. and 5% CO₂. Cells were treated with theircorresponding drugs for 24 hr and then washed with three rinses of PBS(10 ml per rinse) and provided with new media. The cells were thenfurther incubated for colony formation and the colonies were stainedusing a staining solution consisting of: 5 g Crystal Violet, 700 mlmethanol and 300 ml dH₂O. The flasks were then destained with threerinses in cold water. After the third rinse the stained colonies werecounted and the corresponding IC₅₀ for each drug was calculated.

6.22 Example 22 Determination of Radiation Sensitization

To test the ability of the Compounds of the Invention to sensitize cellsto radiation, we used radiation resistant squamous carcinoma cell line,SQ-20B, for initial radiosensitization experiments. SQ-20B is extremelyresistant to ionizing radiation (Do=2.4 Gy in the absence of radiationsensitizers). Briefly, logarithmically growing SQ-20B cells were treatedwith a drug compound at the IC₅₀ concentration (determined using theclonogenic survival assay illustrated in Example 21) for 24 h and thenexposed to graded dose of gamma radiation. Clonogenic survivals weredetermined and fit to the single hit multi-target and the linearquadratic models.

The shape of radiation survival curves are determined by using eitherthe single-hit multitarget model or the linear-quadratic model. Themultitarget model is used to describe the radiation sensitivity of cellsdefined by the terminal slope of the radiation survival curve, which isreferred to as Do. The steeper the slope, the smaller is the value of Doand the more radiation sensitive is the cellular response.Alternatively, a less steep slope results in a larger Do and a moreresistant radiation response. The linear-quadratic model is also used todescribe the radiation sensitivity defined by two components to cellkilling by radiation: one is proportional to dose (αD) and the other isproportional to the square of the dose (βD²). Thus, the dose at whichthe linear and quadratic components are equal is the ratio α/β.

Shown in Table 4 are the results of selected compounds of the inventionin the radiation clonigenic survival assay.

TABLE 4 Radiation Clonigenic Survival Data for Selected CompoundsCompound Do in SQ-20B Compound No. Cells (Gy) α β

TSA 1.65 0.1260 0.03059

SAHA 1.88 0.01197 0.03152

 2 1.62 0.1681 0.03301

 3 1.72 0.2420 0.02486

 5 1.78 0.1978 0.02561

 6 1.94 0.2846 0.01761

 7 1.57 0.1391 0.03335

 8 1.78 0.2378 0.02282

10 2.54 0.1135 0.01558 *For SQ-20B cells in the absence of radiationsensitizers, the Do is 2.4 Gy.

7. REFERENCES CITED

Each and every reference cited herein is hereby incorporated in itsentirety for all purposes to the same extent as if each reference wereindividually incorporated by reference. Furthermore, while the inventionhas been described in detail with reference to preferred embodimentsthereof, it will be apparent to one skilled in the art that variouschanges can be made, and equivalents employed, without departing fromthe scope of the invention.

1. A compound having the formula:

or a pharmaceutically acceptable salt thereof, wherein R¹ is —C₁-C₆alkyl, —C₃-C₇ cycloalkyl, or adamantyl, any of which may beunsubstituted or substituted with one or more of the following groups:—C₁-C₆ alkyl, —O—(C₁-C₆ alkyl), —OH, —CN, —COOR′, —OC(O)R′, NHR′,N(R′)₂, —NHC(O)R′ or —C(O)NHR′ groups wherein R′ is —H or unsubstituted—C₁-C₆ alkyl; m is an integer ranging from 0-10; and n is 4, 5, 6, or 7.2. The compound of claim 1, wherein m is
 0. 3. The compound of claim 1,wherein m is
 1. 4. The compound of claim 1, wherein R¹ is adamantyl. 5.The compound of claim 1, wherein the compound is